U.S. patent application number 12/927040 was filed with the patent office on 2011-04-07 for device for actively removing a target component from blood or lymph of a vertebrate subject.
This patent application is currently assigned to Searete LLC, a limited liability corporation of the State of Delaware. Invention is credited to Roderick A. Hyde, Muriel Y. Ishikawa, Edward K.Y. Jung, Robert Langer, Eric C. Leuthardt, Nathan P. Myhrvold, Elizabeth A. Sweeney, Lowell L. Wood, JR..
Application Number | 20110082412 12/927040 |
Document ID | / |
Family ID | 43535375 |
Filed Date | 2011-04-07 |
United States Patent
Application |
20110082412 |
Kind Code |
A1 |
Hyde; Roderick A. ; et
al. |
April 7, 2011 |
Device for actively removing a target component from blood or lymph
of a vertebrate subject
Abstract
Devices, systems, and methods are disclosed herein for
modulating the levels of one or more target components in the blood
and/or lymph of a vertebrate subject for treatment of a disease or
condition in the vertebrate subject. An implantable device is
provided which includes a body defining at least one lumen
configured for fluid flow; at least one first reservoir in
communication with at least one lumen; one or more energy sources
configured to provide energy to elicit one or more signal responses
associated with one or more labels that bind to one or more target
components in one or more of blood fluid or lymph fluid of the
vertebrate subject; one or more sensors configured to detect the
one or more signal responses associated with one or more labeled
target components; and one or more reactive components in
communication with the at least one lumen for release responsive to
the one or more sensors.
Inventors: |
Hyde; Roderick A.; (Redmond,
WA) ; Ishikawa; Muriel Y.; (Livermore, CA) ;
Jung; Edward K.Y.; (Bellevue, WA) ; Langer;
Robert; (Newton, MA) ; Leuthardt; Eric C.;
(St. Louis, MO) ; Myhrvold; Nathan P.; (Bellevue,
WA) ; Sweeney; Elizabeth A.; (Seattle, WA) ;
Wood, JR.; Lowell L.; (Bellevue, WA) |
Assignee: |
Searete LLC, a limited liability
corporation of the State of Delaware
|
Family ID: |
43535375 |
Appl. No.: |
12/927040 |
Filed: |
November 5, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12803449 |
Jun 25, 2010 |
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12927040 |
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12380399 |
Feb 25, 2009 |
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12803449 |
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12380400 |
Feb 25, 2009 |
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12380399 |
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12660926 |
Mar 5, 2010 |
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12380400 |
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12660928 |
Mar 5, 2010 |
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12660926 |
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Current U.S.
Class: |
604/20 |
Current CPC
Class: |
A61M 5/14276 20130101;
A61M 5/1723 20130101 |
Class at
Publication: |
604/20 |
International
Class: |
A61N 1/00 20060101
A61N001/00 |
Claims
1.-102. (canceled)
103. A method for treating an inflammatory disease or condition in
a vertebrate subject comprising: binding one or more target
components with at least one label in one or more of blood fluid or
lymph fluid of the vertebrate subject; providing energy to elicit
one or more signal responses associated with one or more labeled
target components; detecting the one or more signal responses
associated with the one or more labeled target components; and
providing one or more reactive components to modulate a
physiological effect of the one or more target components in the
one or more of blood fluid or lymph fluid of the vertebrate
subject.
104. The method of claim 103, further including controlling flow of
the one or more of blood fluid or lymph fluid through at least one
lumen with at least one controllable flow barrier.
105. The method of claim 104, further including controlling flow of
the one or more of blood fluid or lymph fluid through the at least
one lumen with at least one controller in communication with one or
more sensors and in communication with the at least one
controllable flow barrier.
106. The method of claim 105, wherein the at least one controller
includes a processor.
107. The method of claim 103, further including providing at least
one first reservoir including the at least one label, wherein the
at least one first reservoir is in communication with a body
including at least one lumen configured for fluid flow, and the at
least one first reservoir is responsive to at least one
controller.
108. The method of claim 105, wherein the at least one controller
is configured to control flow of the one or more of blood fluid or
lymph fluid based on the one or more signal responses associated
with the one or more labeled target components.
109. The method of claim 105, wherein the at least one controller
is configured to control flow of the one or more of blood fluid or
lymph fluid based on the label on the one or more target
components.
110. The method of claim 105, wherein the at least one controller
is configured to control flow of the one or more of blood fluid or
lymph fluid and to control the presence of the one or more reactive
components based on the label on the one or more target
components.
111. The method of claim 103, further including controlling one or
more energy sources with at least one controller in communication
with one or more sensors, wherein the at least one controller is
configured to control the one or more energy sources configured to
provide the energy to elicit the one or more signal responses
associated with the one or more labeled target components.
112. The method of claim 103, further including controlling release
of one or more reactive components with at least one controller in
communication with one or more sensors, wherein the at least one
controller is configured to control release of the one or more
reactive components configured to modulate the physiological effect
of the one or more target components.
113. The method of claim 103, wherein one or more sensors and at
least one controller are configured to control levels of the one or
more target components to substantially attain a target level of
the one or more target components in the one or more of blood fluid
or lymph fluid of the vertebrate subject.
114. The method of claim 113, wherein the one or more sensors and
the at least one controller are configured to control levels of the
detected one or more target components to limit a deviation from
the target level.
115. (canceled)
116. The method of claim 113, wherein the target level includes a
desired concentration of the one or more target components in the
one or more of blood fluid or lymph fluid.
117. The method of claim 113, wherein the target level includes a
desired range of concentrations of the one or more target
components in the one or more of blood fluid or lymph fluid.
118. The method of claim 113, wherein the target level includes a
desired ratio of concentrations of two or more target components in
the one or more of blood fluid or lymph fluid.
119. The method of claim 113, wherein the target level includes a
desired ratio of levels of two or more target components in the one
or more of blood fluid or lymph fluid.
120. (canceled)
121. The method of claim 107, wherein at least one of the at least
one first reservoir includes a first gated mechanism responsive to
one or more sensors and configured to release the one or more
labels into the one or more of blood fluid or lymph fluid of the
vertebrate subject.
122. The method of claim 107, wherein at least one of the at least
one first reservoir includes a first gated mechanism responsive to
one or more sensors and configured to expose the one or more labels
to the one or more of blood fluid or lymph fluid of the vertebrate
subject.
123. The method of claim 107, wherein at least one of the at least
one first reservoir is disposed within the at least one lumen.
124. The method of claim 103, wherein the one or more signal
responses further include one or more signal responses associated
with an interior of at least one lumen.
125. The method of claim 103, wherein the one or more signal
responses are associated with an interior of one or more of a blood
vessel or a lymph vessel of the vertebrate subject.
126. The method of claim 103, wherein the one or more labeled
target components include one or more of circulating cells or
circulating emboli.
127. The method of claim 103, wherein the one or more labeled
target components include one or more of tumor cells, emboli,
misfolded proteins, aggregated proteins, autoimmune antibodies,
infectious agents, or infected cells.
128. The method of claim 103, wherein the one or more labeled
target components include one or more labeled target cells.
129. (canceled)
130. The method of claim 128, wherein one or more sensors are
configured to detect the labeled target cell prior to obtaining a
high resolution image of the labeled target cell.
131. The method of claim 130, wherein the one or more sensors are
configured to correlate the target cell image and the detected
label.
132. (canceled)
133. The method of claim 103, wherein one or more sensors are
external to at least one lumen.
134. The method of claim 103, wherein one or more sensors are
internal to at least one lumen.
135. (canceled)
136. The method of claim 103, further including two or more
parallel lumen configured to receive the one or more target
components.
137. The method of claim 136, wherein a diameter of each of the two
or more parallel lumen is approximately less than two cell
diameters.
138-141. (canceled)
142. The method of claim 103, wherein the one or more reactive
components alters, arrests, disrupts, destroys, inactivates or
ablates the one or more target components.
143. (canceled)
144. The method of claim 103, wherein the one or more reactive
component is configured within at least one second reservoir.
145. (canceled)
146. The method of claim 144, wherein the at least one second
reservoir includes a second gated mechanism responsive to one or
more sensors and configured to release the one or more reactive
components.
147. The method of claim 144, wherein the at least one second
reservoir includes a second gated mechanism responsive to one or
more sensors and configured to expose the one or more reactive
components.
148. The method of claim 144, wherein the one or more reactive
components include one or more reactive chemical components.
149.-150. (canceled)
151. The method of claim 148, wherein the one or more reactive
chemical components are configured to bind to at least one lumen or
are configured to be released into the at least one lumen.
152. The method of claim 148, wherein the one or more reactive
chemical components include one or more of a denaturing agent,
degradative agent, or binding agent.
153.-157. (canceled)
158. The method of claim 103, wherein the one or more reactive
components include one or more reactive biologic components.
159. The method of claim 158, wherein the one or more reactive
biologic components include one or more phagocytic cell types.
160. The method of claim 158, wherein the at least one second
reservoirs include a source for producing the one or more reactive
biologic components.
161. The method of claim 158, wherein the source includes at least
one producer.
162. The method of claim 161, wherein the at least one producer
includes at least one encapsulated cell.
163. (canceled)
164. The method of claim 162, wherein the at least one encapsulated
cell includes at least one genetically-engineered cell.
165.-167. (canceled)
168. The method of claim 158, wherein the one or more reactive
biologic components include a protein, lipid micelle, liposome,
polymer, catalytic antibody, or a combination thereof.
169. (canceled)
170. The method of claim 103, wherein the one or more reactive
components include one or more reactive physical components.
171. The method of claim 170, wherein the one or more reactive
physical components include one or more of polymers, imprinted
polymers, or charged polymers.
172. The method of claim 105, wherein the one or more sensors
include a biosensor, chemical sensor, physical sensor, or optical
sensor.
173. (canceled)
174. The method of claim 172, wherein the one or more sensors
include one or more of a recognition-based substrate, an
aptamer-based substrate, an antibody-based substrate, surface
plasmon resonance, genetically-modified cells, or
genetically-modified cells with receptor-linked signaling.
175. (canceled)
176. The method of claim 105, wherein the one or more sensors are
configured to target a device to a site having an elevated level of
the target components.
177.-180. (canceled)
181. The method of claim 152, wherein the one or more binding
agents are on a matrix adapted to the one or more second
reservoirs, wherein the one or more binding agents are configured
to sequester at least one of the one or more target components from
the one or more of blood fluid or lymph fluid.
182. The method of claim 181, wherein the one or more binding
agents include one or more of antibodies, receptors, or cognates
configured to bind to at least one of the one or more target
components.
183.-186. (canceled)
187. A method for modulating an inflammatory disease or condition
in a vertebrate subject comprising: binding one or more target
components with at least one label in one or more of blood fluid or
lymph fluid of the vertebrate subject; providing energy to elicit
one or more signal responses associated with one or more labeled
target components; detecting the one or more signal responses
associated with the one or more labeled target components; and
providing one or more reactive chemical components to modulate a
physiological effect of the one or more labeled components in the
one or more of blood fluid or lymph fluid of the vertebrate
subject.
188.-189. (canceled)
190. The method of claim 187, further including controlling flow of
the one or more of blood fluid or lymph fluid through at least one
lumen with at least one controllable flow barrier.
191. The method of claim 190, further including controlling flow of
the one or more of blood fluid or lymph fluid through the at least
one lumen with at least one controller in communication with one or
more sensors, and in communication with the at least one
controllable flow barrier.
192.-379. (canceled)
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is related to and claims the benefit
of the earliest available effective filing date(s) from the
following listed application(s) (the "Related Applications") (e.g.,
claims earliest available priority dates for other than provisional
patent applications or claims benefits under 35 USC .sctn.119(e)
for provisional patent applications, for any and all parent,
grandparent, great-grandparent, etc. applications of the Related
Application(s)). All subject matter of the Related Applications and
of any and all parent, grandparent, great-grandparent, etc.
applications of the Related Applications is incorporated herein by
reference to the extent such subject matter is not inconsistent
herewith.
RELATED APPLICATIONS
[0002] For purposes of the USPTO extra-statutory requirements, the
present application constitutes a continuation-in-part of U.S.
patent application Ser. No. 12/380,399, entitled DEVICE, SYSTEM,
AND METHOD FOR CONTROLLABLY REDUCING INFLAMMATORY MEDIATORS IN A
SUBJECT, naming Roderick A. Hyde, Muriel Y. Ishikawa, Edward K. Y.
Jung, Robert Langer, Eric C. Leuthardt, Nathan P. Myhrvold,
Elizabeth A. Sweeney and Lowell L. Wood, Jr. as inventors, filed 25
Feb. 2009, which is currently co-pending, or is an application of
which a currently co-pending application is entitled to the benefit
of the filing date.
[0003] For purposes of the USPTO extra-statutory requirements, the
present application constitutes a continuation-in-part of U.S.
patent application Ser. No. 12/380,400, entitled DEVICE, SYSTEM,
AND METHOD FOR CONTROLLABLY REDUCING INFLAMMATORY MEDIATORS IN A
SUBJECT, naming Roderick A. Hyde, Muriel Y. Ishikawa, Edward K. Y.
Jung, Robert Langer, Eric C. Leuthardt, Nathan P. Myhrvold,
Elizabeth A. Sweeney and Lowell L. Wood, Jr. as inventors, filed 25
Feb. 2009, which is currently co-pending, or is an application of
which a currently co-pending application is entitled to the benefit
of the filing date.
[0004] For purposes of the USPTO extra-statutory requirements, the
present application constitutes a continuation-in-part of U.S.
patent application Ser. No. 12/660,926, entitled DEVICE FOR
PASSIVELY REMOVING A TARGET COMPONENT FROM BLOOD OR LYMPH OF A
VERTEBRATE SUBJECT, naming Roderick A. Hyde, Muriel Y. Ishikawa,
Edward K. Y. Jung, Robert Langer, Eric C. Leuthardt, Nathan P.
Myhrvold, Elizabeth A. Sweeney and Lowell L. Wood, Jr. as
inventors, filed 5 Mar. 2010, which is currently co-pending, or is
an application of which a currently co-pending application is
entitled to the benefit of the filing date.
[0005] For purposes of the USPTO extra-statutory requirements, the
present application constitutes a continuation-in-part of U.S.
patent application Ser. No. 12/660,928, entitled DEVICE FOR
ACTIVELY REMOVING A TARGET CELL FROM BLOOD OR LYMPH OF A VERTEBRATE
SUBJECT, naming Roderick A. Hyde, Muriel Y. Ishikawa, Edward K. Y.
Jung, Robert Langer, Eric C. Leuthardt, Nathan P. Myhrvold,
Elizabeth A. Sweeney and Lowell L. Wood, Jr. as inventors, filed 5
Mar. 2010, which is currently co-pending, or is an application of
which a currently co-pending application is entitled to the benefit
of the filing date.
[0006] The United States Patent Office (USPTO) has published a
notice to the effect that the USPTO's computer programs require
that patent applicants reference both a serial number and indicate
whether an application is a continuation or continuation-in-part.
Stephen G. Kunin, Benefit of Prior-Filed Application, USPTO
Official Gazette Mar. 18, 2003, available at
http://www.uspto.gov/web/offices/com/sol/og/2003/week11/patbene.htm.
The present Applicant Entity (hereinafter "Applicant") has provided
above a specific reference to the application(s) from which
priority is being claimed as recited by statute. Applicant
understands that the statute is unambiguous in its specific
reference language and does not require either a serial number or
any characterization, such as "continuation" or
"continuation-in-part," for claiming priority to U.S. patent
applications. Notwithstanding the foregoing, Applicant understands
that the USPTO's computer programs have certain data entry
requirements, and hence Applicant is designating the present
application as a continuation-in-part of its parent applications as
set forth above, but expressly points out that such designations
are not to be construed in any way as any type of commentary and/or
admission as to whether or not the present application contains any
new matter in addition to the matter of its parent
application(s).
SUMMARY
[0007] Devices, systems, and methods are disclosed herein for
controlling or modulating the levels of one or more target
components in the blood and/or lymph of a vertebrate subject. The
device or system is useful in a method for treating a disease
and/or condition mediated by or indicated by the one or more target
components. The one or more target components include, but are not
limited to, cellular components (e.g., blood cells, cancer cells,
pathogens), non-cellular components (e.g., proteins, lipids,
sugars, carbohydrates, small molecules), or combinations thereof.
An implantable device is provided which includes a body defining at
least one lumen configured for fluid flow; at least one first
reservoir in communication with the at least one lumen; one or more
energy sources configured to provide energy to elicit one or more
signal responses associated with one or more labels that bind to
one or more target components in one or more of blood fluid or
lymph fluid of the vertebrate subject; one or more sensors
configured to detect the one or more signal responses associated
with one or more labeled target components; and one or more
reactive components in communication with the at least one lumen
and for release responsive to the one or more sensors, wherein the
one or more reactive components are configured to modulate a
physiological effect of the one or more target components in the
one or more of blood fluid or lymph fluid of the vertebrate
subject. The one or more reactive components can be configured to
alter, arrest, disrupt, destroy, inactivate, or ablate the one or
more target components.
[0008] Examples of diseases or conditions include, but are not
limited to, acute and chronic inflammatory diseases (e.g., sepsis,
multiple organ dysfunction syndrome, autoimmune disease, asthma,
rhinitis, rheumatoid arthritis), cardiovascular disease,
gastrointestinal disease, cancer, metabolic disease, bacterial
infection (e.g., Staphylococcus bacteremia), viral infection (e.g.,
acquired immunodeficiency syndrome, hepatitis), parasite infection
(e.g., malaria), or chemical or biological agent exposure (e.g.,
drug overdose, environmental toxin).
[0009] In an aspect, the device including the at least one first
reservoir can include the one or more labels. In an aspect, at
least one of the at least one first reservoir can include a first
gated mechanism responsive to the one or more sensors and
configured to release the one or more labels into the one or more
of blood fluid or lymph fluid of the vertebrate subject. At least
one of the at least one first reservoir can include a first gated
mechanism responsive to the one or more sensors and configured to
expose the one or more labels to the one or more of blood fluid or
lymph fluid of the vertebrate subject. At least one of the at least
one first reservoir can be disposed within the at least one lumen.
The one or more signal responses can include one or more of
electromagnetic signal response, optical signal response, visible
light signal response, infrared signal response, signal image
response, fluorescent signal response, radiofrequency signal
response, magnetic signal responses, or surface plamon resonance
response. The one or more signal responses can further include one
or more signal responses associated with an interior of the at
least one lumen. The one or more signal responses can be associated
with an interior of the one or more of a blood vessel or a lymph
vessel of the vertebrate subject. The one or more labeled target
components can include one or more of circulating cells or
circulating emboli. The one or more labeled target components can
include one or more of tumor cells, emboli, misfolded proteins,
aggregated proteins, autoimmune antibodies, infectious agents, or
infected cells. The one or more labeled target components can
include one or more labeled target cells. The one or more labeled
target components can include one or more of cancer cells,
autoimmune-related cells, B cells, T cells, parasites, viruses,
bacteria, fungi, or infected cells. In an aspect, the one or more
labeled target components can include an intracellular target
component. The intracellular target component can be modified by
parasite infection, viral infection, or bacterial infection. The
intracellular target component can be unmodified by an infectious
agent.
[0010] The device including the one or more sensors can be
configured to detect the labeled target cell prior to obtaining a
high resolution image of the labeled target cell. The one or more
sensors can be configured to correlate the target cell image and
the detected label. The one or more sensor can be configured to
function in, or proximal to, the one or more blood vessel or lymph
vessel. The one or more sensor can be external to the at least one
lumen. The one or more sensor can be internal to the at least one
lumen. The one or more labels can include one or more of a
fluorescent label, electromagnetic-emitting label, magnetic label,
electromagnetic label, paramagnetic label, quantum dot label,
aptamer label, gold label, radioactive label,
radiometrically-discernible label, or prodrug.
[0011] In an aspect, the device can include a single unit. In an
aspect, the device can include two or more separate units. In an
aspect, the device can be configured to report to an outside source
or to a computing device, and the device can be configured to
report the detected one or more signal responses associated with
the one or more labeled target components. In an aspect, the device
can further include two or more parallel lumen configured to
receive the one or more target components. In an aspect, a diameter
of each of the two or more parallel lumen can be approximately less
than two cell diameters. The diameter of each of the two or more
parallel lumen can be approximately less than 10 .mu.m. The two or
more parallel lumen can further include 100 or more parallel lumen.
The two or more parallel lumen can further include 1000 or more
parallel lumen. The two or more parallel lumen can further include
10,000 or more parallel lumen. In an aspect, the device is
configured to be placed relative to a tumor or an organ in the
vertebrate subject.
[0012] The device including the at least one second reservoir can
include a second gated mechanism responsive to the one or more
sensors and configured to release the one or more reactive
components. The at least one second reservoir can include a second
gated mechanism responsive to the one or more sensors and
configured to expose the one or more reactive components. The one
or more reactive components can include one or more reactive
chemical components. The one or more reactive chemical components
can include one or more of a binding molecule, antibody, binding
mimetic, synthetic polymer, lectin, integrin, or selectin. The one
or more reactive chemical components can be configured to produce
necrosis or apoptosis in the one or more labeled target components.
The one or more reactive chemical components can include one or
more of a denaturing agent, degradative agent, or binding agent.
The one or more reactive chemical components can be configured to
bind to the at least one lumen or are configured to be released
into the at least one lumen. The one or more binding agents can
include one or more of antibodies, receptors, or cognates
configured to bind to one or more labeled target components. The
one or more binding agents can include one or more of lectin,
binding protein, catalytic antibody, catalytic aptamer, protease,
or photoactivatable agent. The one or more denaturing agents can
include at least of an acid, base, solvent, detergent,
cross-linking agent, chaotropic agent, disulfide bond reducer,
enzyme, drug, cell, or radical ion. The one or more degradative
agents can include at least one of an enzyme, coenzyme, enzyme
complex, catalytic antibody, protease, lipase, proteasomal
component, strong acid, strong base, radical, photoactivatable
agent, drug, cell, or radical ion. The catalytic antibody can
generate a radical ion.
[0013] In an aspect, the device including the one or more reactive
components can include one or more reactive components within at
least one second reservoir disposed within the at least one lumen.
In an aspect, the one or more reactive components can include, but
are not limited to, one or more reactive chemical components, one
or more reactive biologic components, or one or more reactive
physical components. The one or more reactive components can
include one or more reactive biologic components. The one or more
reactive biologic components can include one or more phagocytic
cell types. The one or more second reservoirs can include a source
for producing the one or more reactive biologic components. The
source can include at least one producer. The at least one producer
can include at least one encapsulated cell. The at least one
encapsulated cell can produce the one or more reactive biologic
components. The at least one encapsulated cell can include at least
one genetically engineered cell. The at least one encapsulated cell
can include at least one of a mammalian cell, bacterial cell, yeast
cell, plant cell, insect cell, artificial cell, or enucleated cell.
The at least one encapsulated cell can include one or more of a
myeloid cell; lymphocyte, or precursor thereof. The at least one
encapsulated cell can include one or more of a T-lymphocyte,
B-lymphocyte, macrophage, monocyte, neutrophil, or NK cell. The one
or more reactive biologic components can include a protein, lipid
micelle, liposome, polymer, a catalytic antibody, or a combination
thereof. The catalytic antibody can include a radical ion
generator. The one or more reactive components can include one or
more reactive physical components. The one or more reactive
physical components can include one or more of polymers, imprinted
polymers, or charged polymers.
[0014] In an aspect, the device can further include at least one
controllable flow barrier to the at least one lumen. In an aspect,
the device can further include at least one controller in
communication with the one or more sensors, and in communication
with the at least one controllable flow barrier to the at least one
lumen, wherein the controller is configured to control flow of the
one or more of blood fluid or lymph fluid through the at least one
lumen. The at least one controller can include a processor. The at
least one first reservoir can be responsive to the at least one
controller. The one or more energy sources can be responsive to the
at least one controller. The one or more second reservoirs can be
responsive to the at least one controller. The at least one
controller can be configured to control flow of the one or more of
blood fluid or lymph fluid based on the one or more signal
responses associated with the one or more labeled target
components. The at least one controller can be configured to
control flow of the one or more of blood fluid or lymph fluid based
on the label on the one or more target components. The at least one
controller is configured to control flow of the one or more of
blood fluid or lymph fluid and to control the presence of the one
or more reactive chemical components in the one or more second
reservoirs based on the label on the one or more target
components.
[0015] In an aspect, the device can further include at least one
controller in communication with the one or more sensors, and in
communication with the one or more energy sources, wherein the at
least one controller is configured to control the one or more
energy sources configured to provide energy to elicit one or more
signal responses. In an aspect, the device can further include at
least one controller in communication with the one or more sensors,
wherein the at least one controller is configured to control
release of the one or more reactive components configured to
modulate the physiological effect of the one or more target
components. In an aspect, the device can further include at least
one programmable controller in communication With the at least one
first reservoir.
[0016] In an aspect, the device including the at least one
controller and the one or more sensors can be configured to control
levels of the one or more target components to substantially attain
a target level of the one or more target components in the one or
more of blood fluid or lymph fluid of the vertebrate subject. The
one or more sensors and the at least one controller can be
configured to control levels of the detected one or more target
components to limit a deviation from the target level. The
deviation can be determined by a weighted least squares fit. The
target level can include a desired concentration of the one or more
target components in the one or more of blood fluid or lymph fluid.
The target level can include a desired range of concentrations of
the one or more target components in the one or more of blood fluid
or lymph fluid. The target level can include a desired ratio of
concentrations of two or more target components in the one or more
of blood fluid or lymph fluid. The target level can include a
desired ratio of levels of two or more target components in the one
or more of blood fluid or lymph fluid. The one or more sensors can
include a biosensor, chemical sensor, physical sensor, or optical
sensor. The one or more sensors can include one or more of an
aptamer, antibody, or receptor. The one or more sensors can include
one or more of a recognition-based substrate, an aptamer-based
substrate, an antibody-based substrate, surface plasmon resonance,
genetically modified cells, or genetically modified cells with
receptor-linked signaling. The genetically modified cells can
include receptor-linked signaling by fluorogen-activating proteins.
The one or more sensors can be configured to target the device to a
site having an elevated level of the target components. The one or
more sensors can be configured to detect one or more of cytokines,
T-lymphocytes, B-lymphocytes, antibodies, tumor cells, inflammatory
cells, infected cells, bacteria, parasites, fungi, or viruses. The
one or more sensors can be further configured to detect one or more
of body temperature, vital signs, edema, oxygen level, pathogen
level, or toxin level of the subject. The one or more sensors can
be configured to detect one or more of anaphylatoxin, cytokine,
chemokine, leukotriene, prostaglandin, complement, coagulation
factor, or proinflammatory cytokine. The one or more sensors can be
configured to detect one or more of TNF-.alpha., IL-1, IL-10, IL-6,
IL-8, IL-10, IL-12, LPB, IFN-.gamma., LIF, MIF, MIP-1, MCP-1, C3-a,
C5-a, exotoxin, or endotoxin.
[0017] In an aspect, the device including the one or more binding
agents are on a matrix adapted to the at least one second
reservoirs, wherein the one or more binding agents are configured
to sequester at least one of the one or more target components from
the one or more of blood fluid or lymph fluid. The one or more
binding agents can include one or more of antibodies, receptors, or
cognates configured to bind to at least one of the one or more
target components. The matrix can include one or more of a specific
binding ligand or a hydrophobic surface. The specific binding
ligand or the hydrophobic surface can include one or more of
nucleic acid aptamers, peptide aptamers, molecular imprinting
polymer, antibodies or fragments thereof, high affinity mimetics,
synthetic binding molecules, or receptor binding molecules. The
matrix can include one or more of beads, cells, vesicles, filters,
hydrogel polymers, microparticles, nanoparticles, adsorbent, or
synthetic polymers. The matrix can include one or more of a lectin,
binding protein, receptor, antibody, catalytic antibody, catalytic
aptamer, enzyme, protease conjugate, or photoactivatable agent
conjugate.
[0018] In an aspect, the device is intracorporeal. The device can
be configured to be implanted. The device can include a stent,
bypass implant, nanostructure or microstructure. The device can be
configured to be implanted relative to an organ or tissue in the
subject.
[0019] In an aspect, the device can be at least partially
extracorporeal. The extracorporeal device can include a dialysis
device, hemoperfusion device, apheresis device, intravenous device,
shunt device, or patch device. The device can be tethered to the
blood vessel or the lymph vessel of the vertebrate subject. The
device can be untethered to the blood vessel or the lymph vessel of
the vertebrate subject.
[0020] A method for treating an inflammatory disease or condition
in a vertebrate subject is provided which includes binding one or
more target components with at least one label in one or more of
blood fluid or lymph fluid of the vertebrate subject; providing
energy to elicit one or more signal responses associated with one
or more labeled target components; detecting the one or more signal
responses associated with the one or more labeled target
components; and providing one or more reactive components to
modulate a physiological effect of the one or more target
components in the one or more of blood fluid or lymph fluid of the
vertebrate subject. A method for treating or modulating an
infectious disease or condition in a vertebrate subject is
provided. A method for treating or modulating a neoplastic disease
or condition in a vertebrate subject is provided.
[0021] In an aspect, the method can further include controlling
flow of the one or more of blood fluid or lymph fluid through the
at least one lumen with at least one controllable flow barrier. The
method can further include controlling flow of the one or more of
blood fluid or lymph fluid through the at least one lumen with at
least one controller in communication with one or more sensors and
in communication with the at least one controllable flow barrier.
The at least one controller can include a processor. The method can
further include providing at least one first reservoir including
the at least one label, wherein the at least one first reservoir is
in communication with a body including at least one lumen
configured for fluid flow, and the at least one first reservoir is
responsive to at least one controller. The at least one controller
can be configured to control flow of the one or more of blood fluid
or lymph fluid based on the one or more signal responses associated
with the one or more labeled target components. The at least one
controller can be configured to control flow of the one or more of
blood fluid or lymph fluid based on the label on the one or more
target components. The at least one controller can be configured to
control flow of the one or more of blood fluid or lymph fluid and
to control the presence of the one or more reactive components
based on the label on the one or more target components. The one or
more reactive components can alter, arrest, disrupt, destroy,
inactivate or ablate the one or more target components.
[0022] In an aspect, the method can further include controlling one
or more energy sources with at least one controller in
communication with one or more sensors, wherein the at least one
controller is configured to control the one or more energy sources
configured to provide the energy to elicit the one or more signal
responses associated with the one or more labeled target
components. In an aspect, the method can further include
controlling release of one or more reactive components with at
least one controller in communication with one or more sensors,
wherein the at least one controller is configured to control
release of the one or more reactive components configured to
modulate the physiological effect of the one or more target
components. The one or more sensors and at least one controller can
be configured to control levels of the one or more target
components to substantially attain a target level of the one or
more target components in the one or more of blood fluid or lymph
fluid of the vertebrate subject. The one or more sensors and the at
least one controller can be configured to control levels of the
detected one or more target components to limit a deviation from
the target level. The deviation can be determined by a weighted
least squares fit. The target level can include a desired
concentration of the one or more target components in the one or
more of blood fluid or lymph fluid. The target level can include a
desired range of concentrations of the one or more target
components in the one or more of blood fluid or lymph fluid. The
target level can include a desired ratio of concentrations of two
or more target components in the one or more of blood fluid or
lymph fluid. The target level can include a desired ratio of levels
of two or more target components in the one or more of blood fluid
or lymph fluid.
[0023] In an aspect, the method including the at least one first
reservoir can include the one or more labels. In an aspect, at
least one of the at least one first reservoir can include a first
gated mechanism responsive to the one or more sensors and
configured to release the one or more labels into the one or more
of blood fluid or lymph fluid of the vertebrate subject. In an
aspect, at least one of the at least one first reservoir can
include a first gated mechanism responsive to the one or more
sensors and configured to expose the one or more labels to the one
or more of blood fluid or lymph fluid of the vertebrate subject. In
an aspect, at least one of the at least one first reservoir can be
disposed within the at least one lumen. The one or more signal
responses can further include one or more signal responses
associated with an interior of the at least one lumen. The one or
more signal responses can be associated with an interior of one or
more of a blood vessel or a lymph vessel of the vertebrate subject.
The one or more labeled target components can include one or more
of circulating cells or circulating emboli. The one or more labeled
target components can include one or more of tumor cells, emboli,
misfolded proteins, aggregated proteins, autoimmune antibodies,
infectious agents, or infected cells.
[0024] The method including the one or more labeled target
components can include one or more labeled target cells. The one or
more labeled target components can include cancer cells,
autoimmune-related cells, B cells, T cells, parasites, viruses,
bacteria, fungi, or infected cells. The one or more sensors can be
configured to detect the labeled target cell prior to obtaining a
high resolution image of the labeled target cell. The one or more
sensors can be configured to correlate the target cell image and
the detected label. The one or more sensors can be configured to
function in, or proximal to, the one or more blood vessel or lymph
vessel. The one or more sensors can be external to the at least one
lumen. The one or more sensors can be internal to the at least one
lumen. In an aspect, the one or more labels include a fluorescent
label, electromagnetic-emitting label, magnetic label,
electromagnetic label, paramagnetic label, quantum dot label,
aptamer label, gold label, radioactive label,
radiometrically-discernible label, or prodrug.
[0025] In an aspect, the method can further include two or more
parallel lumen configured to receive the one or more target
components. A diameter of each of the two or more parallel lumen
can be approximately less than two cell diameters. A diameter of
each of the two or more parallel lumen can be approximately less
than 10 .mu.m. The two or more parallel lumen can further include
100 or more parallel lumen. The two or more parallel lumen can
further include 1000 or more parallel lumen. The two or more
parallel lumen can further include 10,000 or more parallel
lumen.
[0026] In an aspect, the method can further include placing a
device including the one or more reactive components relative to a
tumor or an organ in the vertebrate subject. In an aspect, the one
or more reactive component can be configured within at least one
second reservoir. At least one of the at least one second reservoir
can be disposed within at least one lumen. The at least one second
reservoir can include a second gated mechanism responsive to the
one or more sensors and configured to release the one or more
reactive components. The at least one second reservoir can include
a second gated mechanism responsive to the one or more sensors and
configured to expose the one or more reactive components. In an
aspect, the one or more reactive components can include one or more
reactive chemical components. The one or more reactive chemical
components can include one or more of adhesion binding molecule,
antibody, binding mimetic, polymer, lectin, integrin, or selectin.
The one or more reactive chemical components can be configured to
produce necrosis or apoptosis in the one or more labeled target
components. The one or more reactive chemical components can be
configured to bind to at least one lumen or are configured to be
released into the at least one lumen. The one or more reactive
chemical components can include one or more of a denaturing agent,
degradative agent, or binding agent. The one or more binding agents
can include one or more of antibodies, receptors, or cognates
configured to bind to one or more labeled target components. The
one or more binding agents can include one or more of lectin,
binding protein, catalytic antibody, catalytic aptamer, protease,
or photoactivatable agent. The one or more denaturing agents can
include at least of an acid, base, solvent, detergent cross-linking
agent, chaotropic agent, disulfide bond reducer, enzyme, drug,
cell, or radical ion. The one or more degradative agents can
include at least one of an enzyme, coenzyme, enzyme complex,
catalytic antibody, protease, lipase, proteasomal component, strong
acid, strong base, radical, photoactivatable agent, drug, cell, or
radical ion. The catalytic antibody can generate a radical ion.
[0027] In an aspect, the method including the one or more reactive
components can include one or more reactive biologic components.
The one or more reactive biologic components can include one or
more phagocytic cell types. The at least one second reservoirs can
include a source for producing the one or more reactive biologic
components. The source includes at least one producer. The at least
one producer can include at least one encapsulated cell. The at
least one encapsulated cell can produce the one or more reactive
biologic components. The at least one encapsulated cell can include
at least one genetically-engineered cell. The at least one
encapsulated cell can include at least one of a mammalian cell,
bacterial cell, yeast cell, plant cell, insect cell, artificial
cell, or enucleated cell. The at least one encapsulated cell can
include one or more of a myeloid cell, lymphocyte, or precursor
thereof. The at least one encapsulated cell can include one or more
of a T-lymphocyte, B-lymphocyte, macrophage, monocyte, neutrophil,
or NK cell. The one or more reactive biologic components can
include a protein, lipid micelle, liposome, polymer, catalytic
antibody, or a combination thereof. The catalytic antibody can
include a radical ion generator. In an aspect, the one or more
reactive components can include one or more reactive physical
components. The one or more reactive physical components can
include one or more of polymers, imprinted polymers, or charged
polymers.
[0028] In an aspect, the method including the one or more sensors
can include a biosensor, chemical sensor, physical sensor, or
optical sensor. The one or more sensors can include one or more of
an aptamer, antibody, or receptor. The one or more sensors can
include one or more of a recognition-based substrate, an
aptamer-based substrate, an antibody-based substrate, surface
plasmon resonance, genetically-modified cells, or
genetically-modified cells with receptor-linked signaling. The
genetically-modified cells can include receptor-linked signaling by
fluorogen-activating proteins. The one or more sensors can be
configured to target a device to a site having an elevated level of
the target components. The one or more sensors can be configured to
detect one or more of cytokines, T-lymphocytes, B-lymphocytes,
antibodies, tumor cells, inflammatory cells, infected cells,
bacteria, parasites, fungi, or viruses. The one or more sensors can
be configured to detect one or more of body temperature, vital
signs, edema, oxygen level, pathogen level, or toxin level of the
subject. The one or more sensors can be configured to detect one or
more of anaphylatoxin, cytokine, chemokine, leukotriene,
prostaglandin, complement, coagulation factor, or proinflammatory
cytokine. The one or more sensors can be configured to detect one
or more of TNF-.alpha., IL-1, IL-1.beta., IL-6, IL-8, IL-10, IL-12,
LPB, IFN-.gamma., LIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin, or
endotoxin.
[0029] In an aspect, the method including the one or more binding
agents can be on a matrix adapted to the one or more second
reservoirs, wherein the one or more binding agents can be
configured to sequester at least one of the one or more target
components from the one or more of blood fluid or lymph fluid. The
one or more binding agents can include one or more of antibodies,
receptors, or cognates configured to bind to at least one of the
one or more target components. The matrix can include one or more
of a specific binding ligand or a hydrophobic surface. The specific
binding ligand or the hydrophobic surface can include one or more
of nucleic acid aptamers, peptide aptamers, molecular imprinting
polymer, antibodies or fragments thereof, high affinity mimetics,
synthetic binding molecules, or receptor binding molecules. The
matrix can include one or more of beads, cells, vesicles, filters,
hydrogel polymers, microparticles, nanoparticles, adsorbent, or
synthetic polymers. The matrix can include one or more of a lectin,
binding protein, receptor, antibody, catalytic antibody, catalytic
aptamer, protease conjugate, or photoactivatable agent
conjugate.
[0030] A method for modulating an inflammatory disease or condition
in a vertebrate subject is provided which includes binding one or
more target components with at least one label in one or more of
blood fluid or lymph fluid of the vertebrate subject; providing
energy to elicit one or more signal responses associated with one
or more labeled target components; detecting the one or more signal
responses associated with the one or more labeled target
components; and providing one or more reactive chemical components
to modulate a physiological effect of the one or more labeled
components in the one or more of blood fluid or lymph fluid of the
vertebrate subject.
[0031] A method for treating an infectious disease or condition in
a vertebrate subject is provided which includes binding one or more
target components with at least one label in one or more of blood
fluid or lymph fluid of the vertebrate subject; providing energy to
elicit one or more signal responses associated with one or more
labeled target components; detecting the one or more signal
responses associated with the one or more labeled target
components; and providing one or more reactive chemical components
to modulate a physiological effect of the one or more labeled
components in the one or more of blood fluid or lymph fluid of the
vertebrate subject.
[0032] A method for treating a neoplastic disease or condition in a
vertebrate subject is provided which includes binding one or more
target components with at least one label in one or more of blood
fluid or lymph fluid of the vertebrate subject; providing energy to
elicit one or more signal responses associated with one or more
labeled target components; detecting the one or more signal
responses associated with the one or more labeled target
components; and providing one or more reactive chemical components
to modulate a physiological effect of the one or more labeled
components in the one or more of blood fluid or lymph fluid of the
vertebrate subject.
[0033] A system is provided which includes at least one computer
program included on a computer-readable medium for use with at
least one computer system wherein the computer program includes a
plurality of instructions including one or more instructions for
determining at least one treatment of a vertebrate subject through
an implantable device including one or more instructions for
receiving data including data for delivering a body defining at
least one lumen configured for fluid flow; one or more instructions
for receiving data including data from at least one first reservoir
disposed within the at least one lumen; one or more instructions
for receiving data including data from one or more energy sources
configured to provide energy to elicit one or more signal responses
associated with one or more labels that bind to one or more target
components in one or more of blood fluid or lymph fluid of the
vertebrate subject; one or more instructions for receiving data
including data from one or more sensors configured to detect the
one or more signal responses associated with one or more labeled
target components; and one or more instructions for receiving data
including data regarding one or more reactive components disposed
within the at least one lumen for release responsive to the one or
more sensors, wherein the one or more reactive components are
configured to modulate a physiological effect of the one or more
target components in the one or more of blood fluid or lymph fluid
of the vertebrate subject. In an aspect, the system can further
include one or more instructions for receiving data including data
from at least one controllable flow barrier to the at least one
lumen. In an aspect, the system can further include one or more
instructions for receiving data including data from at least one
controller in communication with the one or more sensors, and in
communication with the at least one controllable flow barrier to
the at least one lumen, wherein the controller is configured to
control flow of the one or more of blood fluid or lymph fluid
through the at least one lumen. The one or more reactive components
can include one or more of the one or more reactive components
within at least one second reservoir disposed within the at least
one lumen. The one or more reactive components can include one or
more of one or more reactive chemical components, one or more
reactive biologic components, or one or more reactive physical
components within at least one second reservoir disposed within the
at least one lumen. A device is provided which includes a system
including a signal-bearing medium including, one or more
instructions for treatment of a vertebrate subject through an
implantable device including one or more instructions for receiving
data including data for delivering a body defining at least one
lumen configured for fluid flow; one or more instructions for
receiving data including data from at least one first reservoir
disposed within the at least one lumen; one or more instructions
for receiving data including data from one or more energy sources
configured to provide energy to elicit one or more signal responses
associated with one or more labels that bind to one or more target
components in one or more of blood fluid or lymph fluid of the
vertebrate subject; one or more instructions for receiving data
including data from one or more sensors configured to detect the
one or more signal responses associated with one or more labeled
target components; and one or more instructions for receiving data
including data regarding one or more reactive components disposed
within the at least one lumen for release responsive to the one or
more sensors, wherein the one or more reactive components are
configured to modulate a physiological effect of the one or more
target components in the one or more of blood fluid or lymph fluid
of the vertebrate subject. In an aspect, the device can further
include one or more instructions for receiving data including data
from at least one controllable flow barrier to the at least one
lumen. In an aspect, the device can further include one or more
instructions for receiving data including data from at least one
controller in communication with the one or more sensors, and in
communication with the at least one controllable flow barrier to
the at least one lumen, wherein the controller is configured to
control flow of the one or more of blood fluid or lymph fluid
through the at least one lumen. The one or more reactive components
can include one or more of the one or more reactive components
within at least one second reservoir disposed within the at least
one lumen. The one or more reactive components can include one or
more of one or more reactive chemical components, one or more
reactive biologic components, or one or more reactive physical
components within at least one second reservoir disposed within the
at least one lumen.
[0034] The foregoing summary is illustrative only and is not
intended to be in any way limiting. In addition to the illustrative
aspects, embodiments, and features described above, further
aspects, embodiments, and features will become apparent by
reference to the drawings and the following detailed
description.
BRIEF DESCRIPTION OF THE FIGURES
[0035] FIG. 1 depicts a diagrammatic view of an aspect of an
embodiment of a device.
[0036] FIG. 2 depicts a diagrammatic view of an aspect of an
embodiment of a device.
[0037] FIG. 3 depict a logic flowchart of a method for treating a
disease or condition in a vertebrate subject.
[0038] FIG. 4 depicts a diagrammatic view of an aspect of an
embodiment of a device.
[0039] FIG. 5 depicts a diagrammatic view of an aspect of an
embodiment of a device.
DETAILED DESCRIPTION
[0040] In the following detailed description, reference is made to
the accompanying drawings, which form a part hereof. In the
drawings, similar symbols typically identify similar components,
unless context dictates otherwise. The illustrative embodiments
described in the detailed description, drawings, and claims are not
meant to be limiting. Other embodiments may be utilized, and other
changes may be made, without departing from the spirit or scope of
the subject matter presented here.
[0041] Devices, systems, and methods are disclosed herein for
controlling or modulating the levels of one or more target
components in the blood fluid and/or lymph fluid of a vertebrate
subject. The device or system is useful in a method for treating a
disease and/or condition mediated by or indicated by the one or
more target components. The one or more target components include,
but are not limited to, cellular components (e.g., blood cells,
cancer cells, pathogens), non-cellular components (e.g., proteins,
lipids, sugars, carbohydrates, small molecules), or combinations
thereof. Examples of diseases or conditions include, but are not
limited to, acute and chronic inflammatory diseases (e.g., sepsis,
multiple organ dysfunction syndrome, autoimmune disease, asthma,
rhinitis, rheumatoid arthritis), cardiovascular disease,
gastrointestinal disease, cancer, metabolic disease, bacterial
infection (e.g., Staphylococcus bacteremia), viral infection (e.g.,
acquired immunodeficiency syndrome, hepatitis), parasite infection
(e.g., malaria), chemical or biological agent exposure (e.g., drug
overdose, environmental toxin).
[0042] An implantable device is provided which includes a body
defining at least one lumen configured for fluid flow; at least one
first reservoir in communication with the at least one lumen; one
or more energy sources configured to provide energy to elicit one
or more signal responses associated with one or more labels that
bind to one or more target components in one or more of blood fluid
or lymph fluid of the vertebrate subject; one or more sensors
configured to detect the one or more signal responses associated
with one or more labeled target components; and one or more
reactive components in communication with the at least one lumen
and for release responsive to the one or more sensors, wherein the
one or more reactive components are configured to modulate a
physiological effect of the one or more target components in the
one or more of blood fluid or lymph fluid of the vertebrate
subject. The one or more reactive components can be configured to
alter, arrest, disrupt, destroy, inactivate, or ablate the one or
more target components. In an aspect, the one or more reactive
components can be within at least one second reservoir disposed
within the at least one lumen. In an aspect, the one or more
reactive components can include, but are not limited to, one or
more reactive chemical components, one or more reactive biologic
components, or one or more reactive physical components. The device
can include at least one controllable flow barrier to the at least
one lumen, and at least one controller in communication with the
one or more sensors, and in communication with the at least one
controllable flow barrier to the at least one lumen, wherein the
controller is configured to control flow of the one or more of
blood or lymph through the at least one lumen.
[0043] The device disclosed herein can include a body defining at
least one lumen configured for fluid flow and configured to
function in, or proximal to, one or more of a blood vessel or a
lymph vessel of the vertebrate subject. The device can include one
or more lumen configured to receive the one or more labeled target
components. Alternatively, the device can include two or more
parallel lumen configured to receive the one or more labeled target
components. The phrase "two or more parallel lumen" includes two or
more lumen that are parallel to each other or at least
substantially parallel to each other. In an aspect, a diameter of
each of the two or more parallel lumen is approximately less than
two cell diameters. In an aspect, a diameter of each of the two or
more parallel lumen is approximately less than 10 .mu.m. The one or
more lumens can include a plurality of parallel lumens, e.g., 10 or
more lumens, 100 or more lumens, 1000 or more lumens, or 10,000 or
more lumens, wherein each of the parallel lumens is less than two
cell diameters in cross sectional width. In an aspect, the device
can be a single unit. In an aspect, the device can include two or
more separate units.
[0044] The device can include at least one first reservoir
configured to include one or more labels that bind to one or more
target components in one or more of blood fluid or lymph fluid of
the vertebrate subject. The one or more labels can include one or
more of a fluorescent label, electromagnetic-emitting label,
magnetic label, electromagnetic label, paramagnetic label, quantum
dot label, aptamer label, gold label, radioactive label,
radiometrically-discernible label, or prodrug, or a combination
thereof. The electromagnetic-emitting label can include one or more
labels emitting a specific EM energy frequency or a range of energy
frequencies across the EM spectrum. The one or more labels can
include a contrast agent label, a visible dye, a radiofrequency
label, or a combination thereof. In an aspect, the one or more
labels bind directly to the one or more target components from the
blood or lymph of a vertebrate subject. In an aspect, the one or
more labels include a binding component configured to bind
selectively to one or more target components. The binding component
of the label can include, but is not limited to, antibodies,
antibody fragments, peptides, oligonucleotides, aptamers, protein
nucleic acids, proteins, viruses, viral particles, enzymes,
receptors, bacteria, cells, cell fragments, inorganic molecules,
organic molecules, artificial binding substrates, e.g., formed by
molecular imprinting, or combinations thereof.
[0045] The device can include one or more energy sources configured
to provide energy to elicit one or more signal responses associated
with the one or more labeled target components. The one or more
energy sources include, but are not limited to, electromagnetic
energy, acoustic energy, magnetic energy, electrical energy, or
combinations thereof.
[0046] The device can include one or more sensors configured to
detect one or more signal responses associated with the one or more
labeled target components. The one or more sensors can include, but
are not limited to, one or more of a biosensor, chemical sensor,
physical sensor, or optical sensor. The sensor type depends on the
nature and/or type of label bound to the one or more target
compounds and the nature or type of signal response information
required. Signal response can measure one or more properties of the
one or more labeled target components. Signal response can include
a photograph, size, concentration, granularity, chemical profile,
magnetic property, electrical property, or a characteristic of the
one or more labeled target components. In an aspect, the one or
more signal responses can include one or more signal responses
associated with an interior of the at least one lumen. In an
aspect, the one or more signal responses can include one or more
signal responses associated with the one or more blood vessel or
lymph vessel of the vertebrate subject. The device can be
configured to report to an outside source or to a computing device,
wherein the device can be configured to report the detected one or
more signal responses associated with the one or more labeled
target components.
[0047] The device can include at least one second reservoir
configured to include one or more reactive components responsive to
the one or more sensors, wherein the one or more reactive
components, e.g., one or more reactive chemical components, one or
more reactive biologic components, or one or more reactive physical
components, are configured to modulate a physiological effect of
the one or more target components in the one or more of blood fluid
or lymph fluid of the vertebrate subject. The at least one second
reservoirs can be located within the at least one lumen. The one or
more reactive components can be configured to alter, arrest,
disrupt, destroy, inactivate, or ablate the one or more target
components. For example, when the target component is one or more
labeled target cells, the one or more reactive components can be
configured to produce necrosis or apoptosis in the one or more
labeled target cells. The one or more reactive components can be
configured to bind to the at least one lumen. In an aspect, the one
or more reactive components can include a bifunctional tag. The one
or more reactive components can be configured to be released into
the at least one lumen. The one or more reactive components can
include, but are not limited to, one or more of a binding agent, a
denaturing agent, a degradative agent, a cytotoxic agent, a
cytostatic agent, an apoptotic agent, a chemotherapeutic agent, an
antibody-toxin agent, or a combination thereof. The one or more
binding agents on a matrix adapted to the at least one lumen can be
configured to sequester at least one of the one or more labeled
target components, e.g., one or more target cells, from the blood
or lymph of the vertebrate subject. The one or more binding agents
can include one or more of antibodies, receptors, or cognates
configured to bind to at least one labeled target component. The at
least one labeled target component can be sequestered by the
binding agent prior to treatment with one or more of a second
binding agent, a denaturing agent, a degradative agent, a cytotoxic
agent, a cytostatic agent, an apoptotic agent, a chemotherapeutic,
an antibody-toxin agent, or a combination thereof.
[0048] The device can include a controller in communication with
the one or more sensors, and in communication with the at least one
controllable flow barrier to the at least one lumen, wherein the
controller is configured to control flow of the one or more of
blood or lymph through the at least one lumen. The controller is in
communication with the at least one first reservoir and the at
least one second reservoir wherein the first reservoir including
the one or more labels and the one or more reactive components are
responsive to the controller. The controller is also in
communication with one or more energy sources configured to provide
energy to elicit a signal response, wherein the energy source is
responsive to the controller. The controller is configured to
control flow of the one or more blood or lymph based on the one or
more signal responses associated with the one or more labeled
target components. The controller is configured to control flow of
the one or more of blood or lymph based on the label on the one or
more target components. The controller is configured to control
flow of the one or more of blood or lymph and to control the
presence of the one or more reactive components delivered from the
one or more second reservoirs, wherein controlling reactive
components is based on the presence of the one or more labeled
target components.
[0049] The device can include the controller and the one or more
sensors which can be configured to attain a target level of the one
or more target components in blood or lymph of the vertebrate
subject. In an aspect, the one or more sensors and the controller
can be configured to control levels of the one or more target
components to substantially attain the target level of the one or
more target components in the one or more of blood or lymph of the
vertebrate subject. The target level of one or more target
components can include a desired concentration of the one or more
target components in the blood or lymph. The target level can
include a desired range of concentrations of the one or more target
components in the blood or lymph. The target level can include a
desired ratio of concentrations of two or more target components in
the blood or lymph. The one or more sensors and the controller can
be configured to control levels of the one or more target
components to limit a deviation from the target level. The
deviation can be determined by a weighted least squares fit.
[0050] A method is disclosed herein for treating a disease or
condition in a vertebrate subject. A method is disclosed herein for
modulating a disease or condition in a vertebrate subject. The
disease or condition can include, but is not limited to, an
inflammatory disease or condition, an infectious disease or
condition, or a neoplastic disease or condition. The method can
include binding one or more target components with at least one
label in one or more of blood fluid or lymph fluid of the
vertebrate subject; providing energy to elicit one or more signal
responses associated with one or more labeled target components;
detecting the one or more signal responses associated with the one
or more labeled target components; and providing one or more
reactive components to modulate a physiological effect of the one
or more target components in the one or more of blood fluid or
lymph fluid of the vertebrate subject. The one or more reactive
components can include one or more reactive components within at
least one second reservoir disposed within the at least one lumen.
The one or more reactive components include, but are not limited
to, one or more reactive chemical components, one or more reactive
biologic components, or one or more reactive physical components.
The method can further include controlling flow of the one or more
of blood fluid or lymph fluid through the at least one lumen with
at least one controllable flow barrier. The method can further
include controlling flow of the one or more of blood fluid or lymph
fluid through the at least one lumen with at least one controller
in communication with the one or more sensors and in communication
with the at least one controllable flow barrier to the at least one
lumen.
[0051] Methods are disclosed herein for treating or modulating
diseases or conditions wherein the disease or condition can be
alleviated, modulated, treated, prevented, reduced or eliminated by
one or more reactive component configured to modulate a
physiological effect of the one or more labeled target components
in the blood or lymph of the vertebrate subject. The disease or
condition can include, but are not limited to, cardiovascular
diseases (e.g., ischemic heart disease, inflammatory heart
disease), metabolic diseases (e.g., diabetes), gastrointestinal
diseases (e.g., colitis, Crohn's disease), bacterial infections
(e.g., Staphylococcus bacteremia, anthrax), viral infections (e.g.,
AIDS, hepatitis, hemorrhagic fever), parasitic infections (e.g.,
malaria, sleeping sickness, Chagas disease), metastatic cancer
(e.g., lung, breast, skin, colon, kidney, prostate, pancreas, and
cervix); blood cancers (e.g., leukemia, lymphoma, Hodgkin's
disease, myeloma); chemical or biological agent exposure (e.g.,
drug overdose, poisoning, exposure to environmental toxin).
Additional examples include a number of inflammatory diseases
including, but not limited to, systemic inflammatory response
syndrome, sepsis, septic shock, multiple organ dysfunction
syndrome, ischemia reperfusion, hyperreactive airway disease,
(e.g., asthma, chronic obstructive pulmonary disease, rhinitis,
sinusitis), allergic reaction, anaphylaxis, autoimmune disease,
infectious disease, pulmonary failure, allograft rejection, graft
versus host disease (GVHD), chronic inflammatory disease, psoriatic
arthritis, rheumatoid arthritis.
[0052] A system is disclosed herein which includes at least one
computer program included on a computer-readable medium for use
with at least one computer system wherein the computer program
includes a plurality of instructions including, one or more
instructions for determining at least one treatment of a vertebrate
subject through an implantable device including one or more
instructions for receiving data including data for delivering a
body defining at least one lumen configured for fluid flow; one or
more instructions for receiving data including data from at least
one first reservoir in communication with the at least one lumen;
one or more instructions for receiving data including data from one
or more energy sources configured to provide energy to elicit one
or more signal responses associated with one or more labels that
bind to one or more target components in one or more of blood fluid
or lymph fluid of the vertebrate subject; one or more instructions
for receiving data including data from one or more sensors
configured to detect the one or more signal responses associated
with one or more labeled target components; and one or more
instructions for receiving data including data from one or more
reactive components in communication with the at least one lumen
for release responsive to the one or more sensors, wherein the one
or more reactive components are configured to modulate a
physiological effect of the one or more target, components in the
one or more of blood fluid or lymph fluid of the vertebrate
subject.
[0053] A device is disclosed herein which includes a system
including a signal-bearing medium including, one or more
instructions for treatment of a vertebrate subject through an
implantable device including one or more instructions for receiving
data including data for delivering a body defining at least one
lumen configured for fluid flow; one or more instructions for
receiving data including data from at least one first reservoir in
communication with the at least one lumen; and one or more
instructions for receiving data including data from one or more
energy sources configured to provide energy to elicit one or more
signal responses associated with one or more labels that bind to
one or more target components in one or more of blood fluid or
lymph fluid of the vertebrate subject; one or more instructions for
receiving data including data from one or more sensors configured
to detect the one or more signal responses associated with one or
more labeled target components; and one or more instructions for
receiving data including data regarding one or more reactive
components in communication with the at least one lumen for release
responsive to the one or more sensors, wherein the one or more
reactive components are configured to modulate a physiological
effect of the one or more target components in the one or more of
blood fluid or lymph fluid of the vertebrate subject.
[0054] With reference to the figures, and with reference now to
FIGS. 1, 2, and 3, depicted is an aspect of a device, system, or
method that can serve as an illustrative environment of and/or for
subject matter technologies. The specific devices and methods
described herein are intended as merely illustrative of their more
general counterparts.
[0055] Referring to FIG. 1, depicted is a partial diagrammatic view
of an illustrative embodiment of an implantable device 100
including a body defining at least one lumen 110 configured for
fluid flow; at least one controllable flow barrier 120 to at least
one lumen; at least one first reservoir 130 in communication with
the at least one lumen and configured to include one or more labels
140 that bind to one or more target components 190 from one or more
of blood fluid or lymph fluid 195 of a vertebrate subject; one or
more energy sources 150 configured to provide energy to elicit one
or more signal responses associated with one or more labeled target
components 160; one or more sensors 170 configured to detect the
one or more signal responses associated with the one or more
labeled target components 160; and at least one second reservoir
180 configured to include one or more reactive chemical components
185 responsive to the labeled target components 160 following
detection by the one or more sensors 170, wherein the one or more
reactive chemical components 185 is configured to modulate a
physiological effect of the one or more target components 160 in
the one or more of blood fluid or lymph fluid 195 of the vertebrate
subject. In an aspect, the one or more reactive chemical components
185 can be incorporated onto a surface of the second reservoir 180
or a surface of the lumen 110 of the device. In an aspect, the one
or more reactive chemical components 185 can be released from the
second reservoir 180 as one or more diffusible agents within the
lumen or second reservoir or into the blood or lymph of the
vertebrate subject.
[0056] Referring to FIG. 2, depicted is a partial diagrammatic view
of an illustrative embodiment of calculations of a target value of
one or more target components in an implantable device including a
body defining at least one lumen configured for fluid flow; at
least one first reservoir in communication with at least one lumen;
one or more energy sources configured to provide energy to elicit
one or more signal responses associated with one or more labels
that bind to one or more target components in one or more of blood
fluid or lymph fluid of a vertebrate subject; one or more sensors
configured to detect the one or more signal responses associated
with one or more labeled target components; and one or more
reactive components in communication with at least one lumen for
release responsive to the one or more sensors, wherein the one or
more reactive components are configured to modulate a physiological
effect of the one or more target components in the one or more of
blood fluid or lymph fluid of the vertebrate subject. In an aspect,
the target value can include a desired concentration of the one or
more target components in the peripheral blood, or the target value
can include a desired range of concentrations of the one or more
target components in the peripheral blood. In an aspect, the target
value can include a desired ratio of concentrations of two or more
target components in the peripheral blood. In an aspect, the target
value can be used to determine relative levels of the target
components. The desired ratio of concentrations can be determined
by any method or means, including for example, by a least squares
fit of the concentrations of the two or more target components. For
example, the desired ratio of concentrations can be determined by a
least squares fit of the concentrations of the two or more target
cells at concentrations x.sub.1, x.sub.2, x.sub.3, and x.sub.4 for
a first inflammatory mediator, X, and at concentrations y.sub.1,
y.sub.2, y.sub.3, and y.sub.4 for a second inflammatory mediator,
Y. The least squares can fit to a line or to a two or three
dimensional space indicating the preferred ratio of the two or more
target cells.
[0057] Referring to FIG. 3, depicted is a logic flowchart of a
method 300 for treating an inflammatory disease or inflammatory
condition in a subject. The method 301 includes binding 302 one or
more target components with at least one label in one or more of
blood fluid or lymph fluid of a vertebrate subject; providing
energy to elicit one or more signal responses associated with one
or more labeled target components; detecting the one or more signal
responses associated with the one or more labeled target
components; and providing one or more reactive components to
modulate a physiological effect of the one or more target
components in the one or more of blood fluid or lymph fluid of the
vertebrate subject. The method can include the one or more reactive
components 303 configured within at least one second reservoir. The
one or more reactive components can include one or more reactive
chemical components 304, reactive biologic components 305, or
reactive physical components 306. The method can further include
controlling one or more energy sources 307 with at least one
controller in communication with one or more sensors, wherein the
at least one controller is configured to control the one or more
energy sources configured to provide the energy to elicit the one
or more signal responses associated with the one or more labeled
target components. The method can further include controlling flow
308 of the one or more of blood fluid or lymph fluid through the at
least one lumen with at least one controller in communication with
the one or more sensors and in communication with the at least one
controllable flow barrier. The method can further include
controlling release 309 of one or more reactive components with at
least one controller in communication with one or more sensors,
wherein the at least one controller is configured to control
release of the one or more reactive components configured to
modulate the physiological effect of the one or more target
components.
Device Functioning in or Proximal to Blood and/or Lymph Vessel
[0058] A device is disclosed herein for controlling or modulating
the levels of one or more target components in the blood and/or
lymph of a vertebrate subject and for treating a disease, condition
or infection in the vertebrate subject. The device can include a
body defining at least one lumen configured for fluid flow. The
device can further include at least one controllable flow barrier
to the at least one lumen, and at least one controller in
communication with the one or more sensors, and in communication
with the at least one controllable flow barrier to the at least one
lumen, wherein the controller is configured to control flow of the
one or more of blood or lymph through the at least one lumen.
[0059] The device for controlling or modulating a physiological
effect of the one or more target components associated with a
disease or condition is configured for use in, or proximal to, one
or more blood vessels and/or lymph vessels of a vertebrate subject.
In an aspect, the device can be an intra-vessel sized device, e.g.,
sufficiently small enough in size to be placed in a blood vessel
and/or a lymph vessel while not necessarily obstructing flow
through the vessel. The device can be inserted into a blood vessel
or lymph vessel. The device can be within the vessel in whole or in
part. Configurations for the device include, but are not limited
to, a substantially tubular structure, with one or more lumens in
fluid communication with the blood or lymph vessel of a vertebrate
subject. In an aspect, the device can take the form of a short
cylinder, an annulus, a cylinder, and/or a spiral. See, e.g., U.S.
Patent Applications 2007/0066929 and 2008/0058785; Bezrouk et al,
Scripta Medica (BRNO) 78(4):219-226, 2005, each of which is
incorporated herein by reference. In an aspect, the device has a
cylindrical and hollow configuration, with a single central
opening, optionally allowing the exterior of the cylindrical
structure to contact and engage the wall of the vessel, and the
interior of the structure (within the single central opening) to
form a fluid-contacting portion of the device. For example, the
device can be configured as a specialized stent fixed within a
specific artery or vein. See, e.g., U.S. Pat. Nos. 5,411,551;
7,326,240; 6,743,190; 6,793,642; 6,488,704; 7,244,232; U.S. Patent
Applications 2007/0294150; 2008/0281400; 2006/01832223;
2008/0286278; Yokota, et al., 22nd IEEE International Conference
MicroElectro Mechanical Systems, Sorrento, Italy, January 25-29.
IEEE pp. 495-499, 2009, each of which are incorporated herein by
reference. For example, in a process for identifying and treating
diseased cells, cells within a living organism can be labeled with
a fluorescent marker in an implanted reservoir. Thereafter, the
labeled cells are allowed to circulate within the organism and
thereafter can be detected with the use of an implanted detector
and either isolated or ablated. See, e.g., U.S. Patent Application
2007/0276208, each of which are incorporated herein by
reference.
[0060] In an aspect, the device can include two or more parallel
lumens configured to receive the one or more target components. The
two or more parallel lumens can be parallel to each other. The two
or more parallel lumens can be parallel to the blood vessel or
lymph vessel of the subject. In an aspect, a diameter of each of
the two or more parallel lumens is approximately less than ten
cells diameters, less than eight cell diameter, less than six cell
diameters, less than four cell diameters, less than two cell
diameters. In an aspect, the diameter of each of the two or more
parallel lumens can be the same diameter. In an aspect, the
diameter of each of the two or more parallel lumens can vary from
lumen to lumen. In an aspect, the diameter of each of the two or
more parallel lumens can vary within a single lumen, e.g., tapering
from wider to narrower. Tapering of the lumen diameter can function
to slow the flow of the one or more target components through the
lumen and increase contact with the one or more sensors and one or
more reactive components. In an aspect, a diameter of each of the
two or more parallel lumen is approximately less than 10 .mu.m. The
at least one lumens can include a plurality of parallel lumens,
e.g., 10 or more lumens, 100 or more lumens, 1000 or more lumens,
or 10,000 or more lumens, wherein each of the parallel lumens is
approximately less than ten cells diameters, less than eight cell
diameter, less than six cell diameters, less than four cell
diameters, or less than two cell diameters in cross sectional
width.
[0061] In an aspect, the device can be configured to be
approximately hemi-spherical or hemi-elliptoid, in whole or in
part, allowing a portion of a cross-section of at least a portion
of the device to contact and/or engage the internal wall of a blood
or lymph vessel without significantly and/or substantially
obstructing the movement of fluid within the vessel. The device can
include one or more wall engaging components including, but not
limited to, rotating wheels, projections (e.g. arms), springs,
hooks (e.g. claws), suction cups, and/or tissue adhesives that are
configured to engage wall portions.
[0062] In an aspect, the device can be configured in a pill- or
capsule-shape, and configured to move through a central portion of
a vessel. The device can engage a wall of the vessel using one or
more engaging components and/or freely travel through the blood
and/or lymph systems. See, e.g., U.S. Patent Application
2007/0156211 A1, which is incorporated herein by reference. The
device can be targeted to a site of disease, e.g., inflammation,
infection, or cancer, in the vertebrate subject. In an aspect, the
device can sense elevated levels of one or more target cells in the
blood or lymphatic system of the vertebrate subject and can target
and form a stationary location at, or near, a site of disease or
condition in the circulation of the vertebrate subject. In an
aspect, the implantable device can be incorporated into a shunt,
e.g., an arteriovenous shunt inserted between an artery and a
vein.
[0063] In an aspect, the device can be positioned, in whole or in
part, proximal to a blood vessel or lymph vessel. "Proximal to" can
refer to a space or area near to a blood vessel or lymph vessel.
Locations that are proximal to a vessel can include, for example,
locations external to the vessel wall where there is space for
implanting one or more devices in whole or in part, and optionally
to facilitate external access to the devices in whole or in part.
In an aspect, "proximal to" can include distances including, but
not limited to, approximately 0.1, 1.0, 10, and/or 100 .mu.m and/or
approximately 0.1, 1.0, 10, and/or 100 mm, and can optionally
include larger and/or smaller distances depending on, for example,
the availability of space and the size of the device and/or the
vessel.
[0064] In an aspect, the device can be configured as a
self-contained unit that includes all functionalities necessary for
operation of the device. In another aspect, the system can be
configured as two or more components in two or more locations
separate from one another, wherein one or more of the components
include one or more essential and/or non-essential functionalities.
As an example, one component of the system can be placed within a
blood vessel, and another component of the system placed proximal
to the blood vessel optionally in a location more accessible from
the exterior of the vertebrate subject, or where there is
additional space. A remote portion can be configured to provide for
monitoring of the vessel portion of the system, data collection, or
data analysis, and/or remote-control of one or more other functions
of the system such as sensing target cells, controlling flow
through a flow route, and releasing a reactive component. The
remote portion can be at a separate location within the body of the
vertebrate subject, or outside the body of the vertebrate subject.
Data and/or power signals can be transmitted between the one or
more components of the device using electromagnetic signals, or
electrical or optical links.
[0065] The dimensions and mechanical properties, e.g., rigidity, of
the device, in whole or in part, can be configured for
compatibility with the location of use in order to provide for
reliable positioning and/or to provide for movement of the device
while preventing damage to the vessel, the vessel lumen, and/or
internal location and its surrounding structure. The choice of
structural component size and configuration appropriate for a
particular blood or lymph vessel location can be selected by a
person of skill in the art. Structural components of the device can
be constructed using a variety of manufacturing methods, from a
variety of biocompatible materials. Appropriate materials include
metals, ceramics, polymers, and composite materials having suitable
biocompatibility, sterilizability, mechanical, and physical
properties. Examples of materials and selection criteria are
described, for example, in The Biomedical Engineering Handbook
(Second Edition, Volume I, J. D. Bronzino, Ed., Copyright 2000, CRC
Press LLC, pp. IV-1-43-22), which is incorporated herein by
reference. Manufacturing techniques can include, but are not
limited to, injection molding, extrusion, die-cutting,
rapid-prototyping, and will depend on the choice of material and
device size and configuration. Sensing and energy-emitting portions
of the devices as well as associated control circuitry can be
fabricated on the structural elements using various
microfabrication and/or MEMS techniques or can be constructed
separately and subsequently assembled to the structural elements,
as one or more distinct components. See, e.g., U.S. Patent
Applications 2005/0221529, 2005/0121411, 2005/0126916,
2007/0066939, 2007/0225633 and Nyitrai, et al. "Preparing Stents
with Masking & Etching Technology" 26.sup.th International
Spring Seminar on Electronics Technology pp. 321-324, IEEE, 2003,
each of which is incorporated herein by reference.
[0066] In additional to biocompatible materials described and
incorporated herein, flexible material having adjustable diameter,
taper, and length properties can be used as part of the structural
material. For example, some materials can change from a longer,
narrower configuration, to a shorter, wider configuration, or can
taper over their length, e.g., shape-memory polymers that can move
from one shape to another in response to a stimulus such as heat.
Structural elements that can exhibit this type of
expansion/contraction property can include self-expanding material,
resilient material, and/or mesh structures formed of various
metals, e.g., ionic polymer-metal composites (IPMC) or plastics,
and some polymeric materials, e.g., hydrogels, nitinol, or
polyester. See, e.g. Bellin et al., Proc. Natl. Acad. Sci. USA.
103:18043-18047, 2006; and Shanpoor et al., Smart Mater. Struct.
14:197-214, 2005, each of which is incorporated herein by
reference.
Device for Controlling or Modulating Levels of One or More Target
Components
[0067] Devices, systems, and methods are disclosed herein for
controlling or modulating the levels of one or more target
components in the blood and/or lymph of a vertebrate subject, and
methods are disclosed for treating a disease, condition or
infection in the vertebrate subject. The target component can be a
normal component or an abnormal component of the blood or lymph of
the vertebrate subject. The target component can be associated with
a normal physiological state or with a pathological state of the
vertebrate subject. The target component can be a non-cellular
component or a cellular component of the blood or lymph of the
vertebrate subject. Examples of non-cellular target components
include, but are not limited to, proteins, lipids, sugars,
minerals, vitamins or combinations thereof. Examples of cellular
target components include one or more circulating cells, e.g., red
blood cells, white blood cells, pathogens, pathogen-infected blood
cells, cancer cells. In an aspect, the one or more target
components can be one or more of a circulating emboli, a misfolded
protein, an aggregated protein, an autoimmune antibody, an
infectious agent, an infected cell, a thrombus, a plaque, a lipid,
an aggregate, a cell, a specific cell type, a cell fragment, a
cellular component, an intracellular component, an organelle, a
collection or aggregation of cells, a cell membrane, a prion, a
therapeutic agent, an illicit drug, a drug of abuse, or a toxin.
See, e.g., He, et al., Proc. Natl. Acad. Sci. USA 104: 11760-11765,
2007, which is incorporated herein by reference. When the target
component is an autoimmune antibody, the reactive component can
include a component of cellular tissue bound by the autoimmune
antibody configured to target the autoimmune antibody.
[0068] In an aspect, the one or more target components are cells
circulating in the blood and/or lymph of a vertebrate subject.
Cellular target components can include, but are not limited to,
blood cells, e.g., platelets, red blood cells, neutrophils,
lymphocytes, monocytes, eosinophils, basophils; or pathogens, e.g.,
viral-infected cells, bacteria, fungus, parasite; or cancer cells,
e.g., metastatic cancer cells, blood cancer cells.
[0069] The one or more cellular target components can include, but
are not limited to, one or more blood cells associated with a
pathological state in which the normal circulating levels of one or
more class of blood cells is elevated in the blood and/or the lymph
of the vertebrate subject. For example, elevated levels of red
blood cells can be associated with exposure to carbon monoxide,
long-term lung disease, kidney disease, cancer, certain forms of
heart disease, or liver disease. Elevated levels of platelets can
be associated with bleeding, iron deficiency, cancer, or bone
marrow pathologies. Elevated levels of neutrophils and eosinophils
can be associated with infection, malignancy or autoimmune
diseases. In an aspect, the cellular target components include
blood cells that are modified or altered as a result of a disease
and/or condition. For example, hyperactivated B-lymphocytes in
patients with inflammatory bowel disease exhibit increased surface
expression of toll-like receptor 2 (TLR2) relative to B-lymphocytes
from normal individuals. See, e.g., Noronha, et al., J. Leukoc.
Biol. 86: Epub ahead of print; Rea, WebMD. Complete Blood Count
(CBC). at www.webmd.com/a-to-z-guides/complete-blood-count-cbc.
Last updated Sep. 12, 2008; Accessed Oct. 5, 2009; each of which is
incorporated herein by reference.
[0070] The one or more cellular target components can include one
or more pathogens circulating in the blood and/or the lymph of the
vertebrate subject. Examples of blood borne pathogens include, but
are not limited to, pathogen-infected cells, e.g., human
immunodeficiency virus (HIV), and the hepatitis B, hepatitis C, and
hepatitis D viruses; bacteria, e.g., Staphylococcus, Streptococcus,
Pseudomonas, Haemophilus, Escherichia coli; fungi, e.g.,
Aspergillus, Candida albicans, Candida glabrata, Torulopsis
glabrata, Candida tropicalis, Candida krusei, and Candida
parapsilosis; and parasites, e.g., Trypanosoma cruzi, Trypanosoma
brucei, Leishmania, Plasmodium, Babesia microti, Toxoplasma gondii.
Other bacterial pathogens that might be found in the blood and/or
lymph at some point during a bacterial infection include, but are
not limited to, Bartonella, Coxiella burnetii, Chlamydia,
Salmonella, Shigella, Yersinia, Legionella, Neisseria,
Mycobacterium tuberculosis, Listeria, Corynebacterium diphtheriae,
Campylobacter, Enterobacter. Other viral-infected cells that can be
found in the blood and/or lymph at some point during a viral
infection include, but are not limited to, cytomegalovirus,
influenza virus, human T-lymphotrophic virus, Epstein-Barr virus,
roseolovirus, herpes lymphotropic virus, Karposi's
sarcoma-associated herpes virus, herpes simplex virus, Ebola virus,
or Marburg virus.
[0071] In another aspect, the one or more cellular target
components can be one or more pathogens or pathogen-infected cells
circulating in the blood and/or the lymph of a vertebrate subject.
The one or more cellular target components can include one or more
circulating blood cells infected with a pathogen including, but not
limited to, bacteria, virus, or parasite. In an aspect, the one or
more cellular target components include circulating blood cells
infected with bacteria, e.g., red blood cells infected with B.
bacilliformis or Bartonella spp. See, e.g., Dehio Cell. Microbiol.
10:1591-1598, 2008; Chomel et al., Vet. Res. 40: 29, 2009, each of
which is incorporated herein by reference. In another aspect, the
one or more cellular target components include one or more cells
infected with HIV, primarily CD4.sup.+ T lymphocytes but also
including macrophages and dendritic cells. In an aspect, the one or
more cellular target components include red blood cells infected
with the malaria parasite Plasmodium falciparum. Red blood cells
infected with P. falciparum can be distinguished from normal red
blood cells by visual inspection, changes in granularities and
changes in surface protein expression including expression on the
red blood cell surface of the parasite derived protein P.
falciparum erythrocyte membrane protein (PfEMP1). See, e.g.,
Dempster & Di Ruperto, Circuits and Systems, 2001. ISCAS 2001.
The 2001 IEEE International Symposium, 5:291-294, 2001; Weatherall,
et al., Hematology Am. Soc. Hematol. Educ. Program 35-57, 2002;
Horata, et al., Malaria J. 8:184, 2009, each of which is
incorporated herein by reference.
[0072] The one or more cellular target components can include one
or more cancer cells circulating in the blood and/or lymph of a
vertebrate subject. In an aspect, the cancer cells can include
circulating tumor cells that have metastasized from solid tumors
located elsewhere in the body. Examples of solid tumors from which
metastatic cells can arise include, but are not limited to,
carcinomas (e.g., adrenal, breast, cervical, colon, endometrial,
lung, ovarian, pancreatic, prostate, stomach, testicular, thyroid,
melanoma, head & neck) and sarcomas (e.g., brain, Ewing's
sarcoma, Karposi's sarcoma, osteosarcoma, spinal cord). Circulating
tumor cells are indicative of metastasis and can suggest a need for
changes in the treatment regime. For example, the detection of
circulating tumor cells in melanoma patients who are clinically
"disease-free" indicates disease recurrence, tumor cell spreading,
and a high potential for distant metastasis, and enables
identification of high-risk melanoma patients. See, e.g., Schuster
et al., Clin. Cancer Res. 13:1171-1178, 2007, which is incorporated
herein by reference. The appearance of circulating tumor cells can
also provide an indication of the long term prognosis for the
patient. For example, breast cancer patients with levels of
circulating tumor cells equal to or higher than five cells per 7.5
milliliters of blood have a shorter median progression-free
survival (2.7 months vs. 7.0 months) and shorter overall survival
(10.1 months versus greater than 18.0 months) as compared with
breast cancer patients with less than five cells per 7.5
milliliters of blood. See, Cristofanilli et al. N. Engl. J. Med.
351:781-791, 2004, which is incorporated herein by reference.
[0073] In another aspect, the cancer cells can be associated with
blood cancers. Examples of blood cancers include, but are not
limited to, lymphoma, various types of leukemia, and multiple
myeloma. Lymphoma is a cancer of lymphocytes which usually begins
in a lymph node but can originate from the stomach, intestines,
skin or any other organ. The two main types of lymphoma are
Hodgkin's disease and non-Hodgkin's lymphoma. In Hodgkin's disease,
the abnormal cells are called the Reed-Sternberg cells, giant
binucleated or multinucleated macrophages. This type of cancer can
spread throughout the lymphatic system, affecting any organ or
lymph tissue in the body. Non-Hodgkin's lymphoma is classified by
the size, type and distribution of cancer cells in the lymph nodes.
Low-grade lymphomas include small-lymphocytic lymphoma, follicular
small-cleaved-cell lymphoma, and follicular mixed-cell lymphoma.
Intermediate-grade lymphomas include follicular large-cell
lymphoma, diffuse small-cleaved-cell lymphoma, diffuse mixed
lymphoma, and diffuse large-cell lymphoma. High-grade lymphomas
include immunoblastic lymphoma, lymphoblastic lymphoma, and small
noncleaved (Burkitt's and non-Burkitt's) lymphoma. Multiple myeloma
is cancer of the bone marrow caused by the uncontrolled growth of
effector B cells. Effector B cells normally make antibodies (e.g.,
immunoglobulins) to fight infections. In multiple myeloma effector
B cells multiply uncontrollably, generating too much of a single
type of immunoglobulin. The level of other immunoglobulins drops,
leaving the patient vulnerable to infection. The cancerous plasma
cells collect in the bones and bone marrow and can form tumors that
destroy the bone tissue, causing the bones to become fragile and
prone to fracture.
[0074] In an aspect, the one or more target components include
non-cellular components present in the blood and/or lymph of a
vertebrate subject. Non-cellular components can include, but are
not limited to, sugars (e.g., glucose), lipids (e.g.,
triacylglycerols, cholesterol, phospholipids), vitamins, minerals,
non-protein hormones (e.g., estrogen, testosterone), proteins
(e.g., enzymes, hormones, antibodies, blood clotting factors,
lipoproteins). Additional examples of proteins found in the blood
and/or lymph include, but are not limited to, serum proteins (e.g.,
subclasses of immunoglobulins, complement factors, Cl esterase,
circulating immune complexes, albumin, anti-trypsin, fetoprotein,
acid glycoprotein, alpha-macroglobulin, beta-microglobulin,
ceruloplasmin, transferrin), acute phase proteins associated with
disease (e.g., C-reactive protein, SPLA2, ferritin), coagulation or
complement related proteins (e.g., tissue-factor pathway inhibitor,
soluble tissue factor, kallikrein, factor XIIa, thrombin, lupus
anticoagulant, soluble CD46, soluble CD55), and markers of cellular
activation (e.g., elastase, elastase/antitrypsin complexes,
lactoferrin, granzym, nucleosomes, soluble CD16, soluble CD27).
[0075] The one or more target components can include one or more
inflammatory mediators. Examples of inflammatory mediators include,
but are not limited to, interferons (IFN) IFN-.alpha., IFN-.beta.,
and IFN-.gamma.; interleukins (IL) IL-1, IL-2, IL-3, IL-4, IL-5,
IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15,
IL-16, IL-17, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-24,
IL-27, IL-28, IL-29, IL-30, IL-31, and IL-32; tumor necrosis factor
(TNF) TNF-.alpha. and TNF-.beta.; granulocyte colony stimulating
factor (G-CSF); granulocyte-macrophage colony stimulating factor
(GM-CSF); macrophage colony-stimulating factor (M-CSF); gelsolin,
erythropoietin (EPO); and thrombopoietin (TPO). The one or more
inflammatory mediators can be any of a number of chemotactic
cytokines (chemokines) including, but not limited to, CC chemokines
CCL1 through CCL28 exemplified by RANTES (CCL5), MCP-1 (CCL2), LARC
(CCL20), MIP-1.alpha. (CCL3), and MDC (CCL22); CXC chemokines CXCL1
through CXCL17 exemplified by LIX (CXCL5), GCP-2 (CXCL6) and BCA-1
(CXCL13); C chemokines XCL1 and XCL2; CX3C chemokine C3CL1
(fractalkine); and chemokine like molecules exemplified by MIF.
Other inflammatory mediators include, but are not limited to,
anaphylatoxin fragments C3a, C4a, and C5a from the complement
pathway; leukotrienes LTA4, LTB4, LTC4, LTD4, LTE4, and LTF4;
prostaglandins; growth factors EGF, FGF-9, FGF-basic, growth
hormone, stem cell factor (SCF), TGF-.beta. and VEGF; soluble
receptors to tumor necrosis factor receptor (sTNFr); soluble
interleukin receptors sIL-1r and sIL-2r; C-reactive protein; CD11b;
histamine; serotonin; apolipoprotein A1; .beta.2-microglobulin;
bradykinin; D-dimer; endothelin-1; eotaxin; factor VII; fibrinogen;
GST; haptoglobin; IgA; insulin; IP-10; leptin; LIF; lymphotactin;
myoglobin; OSM; SGOT; TIMP-1; tissue factor; VCAM-1; VWF;
thromboxane; platelet activating factor (PAF); immunoglobulins; and
endotoxins such as lipopolysaccharide (LPS); and various exotoxins
such as superantigens, e.g., from, Staphylococcus aureus and
Streptococcus pyogenes.
[0076] In an aspect, the one or more target components are
exogenous chemical or biological agents that have been introduced
into the blood and/or lymph of a vertebrate subject. Examples of
exogenous target components include, but are not limited to, drugs,
both legal and illegal, poisons, and environmental toxins. Examples
of drugs commonly used to treat disease include, but are not
limited to, anti-depressants, anti-psychotics, anti-virals,
anti-fungals, anti-parasitics, anti-protozoal drugs,
anti-inflammatory, antibiotics, analgesics, anti-hypertensives,
statins, other cardiovascular drugs, antiseizure drugs, muscle
relaxants, hormones, steroids, chemotherapeutic agents. Examples of
common illicit drugs or drugs of abuse include, but are not limited
to, cannabinoids such as hashish and marijuana; depressants such as
barbiturates, benzodiazepines (e.g., Valium, Halcion), gamma
hydroxy butyrate (GHB), and methaqualone; dissociative anesthetics
such as ketamine and phencyclidine (PCP); hallucinogens such as
LSD, mescaline, ibogaine, and psilocybin; opioids and morphine
derivatives such as codeine, fentanyl, heroin, morphine, opium,
oxycodone (OxyContin) and hydrocodone bitartate/acetaminophen
(Vicodin); stimulants such as amphetamines, methamphetamine,
cocaine, methylphenidate (Ritalin), MDMA (ecstasy), and nicotine;
and anabolic steroids (see "Commonly Abused Drugs", National
Institute on Drug Abuse, www.drugabuse.gov). Examples of
environmental toxins include, but are not limited to, lead,
arsenic, mercury, phthalates. Examples of additional environmental
toxins can be found in ATSDR: Safeguarding Communities from
Chemical Exposures, Centers for Disease Control, and in the Agency
for Toxic Substances & Disease Registry as part of the Centers
for Disease Control (www.atsdr.cdc.gov; Patel et al., J. Med.
Toxicol. 2:83-84, 2006, each of which is incorporated herein by
reference).
Device Including Labels for Labeling Target Components
[0077] The device disclosed herein includes one or more detectable
labels that can target and bind to one or more target components in
the blood or lymph of a vertebrate subject. The one or more
detectable labels include, but are not limited to, at least one of
a fluorescent label, an electromagnetic-emitting label, a quantum
dot label, a gold label, dye, chemiluminescent dye, a prodrug, or a
combination thereof. The one or more detectable labels can include
at least one of a radioactive label; a radiopaque dye; a
radiofrequency identification tag; fluorescent label; chromogenic
label; a contrast agent label, a visible dye, volatile label; mass
label; luminescent label, e.g., bioluminescent or chemiluminescent;
metallic label, e.g., gold particles, magnetic beads, or
paramagnetic beads; dyes, e.g., direct, indirect, or releasable; or
a combination thereof.
[0078] The device including one or more labels configured to bind
to one or more target components can be detected as a
light-emitting label that either naturally emits light or emits
light in response to an excitation energy. An example of a
light-emitting label includes a variety of quantum dots or
semiconductor nanocrystals that fluoresce at various wavelengths in
response to an excitation energy (see, e.g., Jaiswal et al., Nature
Biotech. 21:47-51, 2003, which is incorporated herein by
reference). Examples of other fluorescing dyes for use with
biological samples include, but are not limited to, fluorescein
(FITC), indocyanine green (ICG) and rhodamine B, red and near
infrared emitting fluorophores (600-1200 nm) including cyanine dyes
such as Cy5, Cy5.5, and Cy7 (Amersham Biosciences, Piscataway,
N.J., USA) and/or a variety of Alexa Fluor dyes such as Alexa Fluor
633, Alexa Fluor 635, Alexa Fluor 647, Alexa Fluor 660, Alexa Fluor
680, Alexa Fluor 700 and Alexa Fluor 750 (Molecular
Probes-Invitrogen, Carlsbad, Calif., USA; see, e.g., U.S. Pat. App.
No. 2005/0171434 A1, which is incorporated herein by reference).
Additional fluorophores include IRDye800, IRDye700, and IRDye680
(LI-COR, Lincoln, Nebr., USA), NIR-1 and 105-OSu (Dejindo,
Kumamotot, Japan), LaJolla Blue (Diatron, Miami, Fla., USA),
FAR-Blue, FAR-Green One, and FAR-Green Two (Innosense, Giacosa,
Italy), ADS 790-NS and ADS 821-NS (American Dye Source, Montreal,
Calif.), NIAD-4 (ICx Technologies, Arlington, Va.). Other
fluorescing agents include BODIPY-FL, europium, green, yellow and
red fluorescent proteins, luciferase. Alternatively, the light
emitting marker can be a fluorescently labeled microsphere.
Examples of fluorescently labeled microspheres ranging in diameter
from 20 nanometers to 10 micrometers are available from commercial
sources (e.g., Fluor Spheres.RTM. Fluorescent Microspheres,
Invitrogen, Carlsbad, Calif.).
[0079] The device including one or more labels configured to bind
to one or more target components can be one or more detectable
labels including a radioactive marker. Radioactive labels can
include one or more radioisotope commonly used in nuclear medicine,
including, but are not limited to, iodine-131, cobalt-60,
cesium-137, technetium-99m, carbon-11, nitrogen-13, oxygen-15, and
fluorine-18. Other medical radioisotopes include, but are not
limited to, americium-241, arsenic-74, gold-198, boron-11,
carbon-14, calcium-48, cerium-141, cobalt-55, cobalt-57,
chromium-51, cesium-130, cesium-131, copper-61, copper-62,
copper-64, copper-67, dysprosium-165, europium-155, gallium-67,
gallium-68, gadolinium-153, germanium-68, hydrogen-3, iodine-122,
iodine-123, iodine-124, iodine-125, iodine-132, indium-111,
indium-115m, Iridium-191m, krypton-81m, manganese-51, manganese-52,
Nb-95, osmium-194, phosphorous-32, phosphorous-33, lead-203,
lead-82, ruthenium-97, ruthenium-103, sulfur-35, scandium-46,
selenium-72, selenium-75, strontium-85, tantalum-178, tantalum-182,
terbium-149, thallium-201, xenon-127, xenon-133.
[0080] In an aspect, the device including one or more labels
configured to bind to one or more target components can include a
radioactive marker such as one used for diagnostic positron
emission tomography (PET), single photon emission computed
tomography (SPECT), and gamma camera imaging. Radioisotopes
commonly used for PET, SPECT and gamma camera imaging include, but
are not limited to, fluorine-18, carbon-11, nitrogen-13, oxygen-15,
and fluorine-18; salts of radioisotopes such as I-131 sodium
iodide, Tl-201 thallous chloride, Sr-89 strontium chloride;
technetium Tc-99m; compounds containing iodine-123, iodine-124,
iodine-125, and iodine-131; compounds containing indium-111 such as
.sup.111In-1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic
acid and .sup.111In-Diethylenetriamine pentaacetic acid;
.sup.177Lu-[(R)-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyc-
lohexane-1,2-diamine-pentaacetic acid) (.sup.177Lu-CHX-A''-DTPA),
.sup.64Cu-DOTA, .sup.89Zr, .sup.86Y-DOTA. For example, tritiated
diisopropylfluorophosphate (.sup.3H-DFP) can be used to selectively
label granulocytes of neutrophils. See, e.g., Price et al. Blood
88:335-340, 1996, which is incorporated herein by reference.
[0081] In an aspect, the device including one or more labels
configured to bind to one or more target components can include a
magnetic marker, e.g., magnetic beads, magnetic particles or carbon
nanotubes. Magnetic beads and magnetic particles of various
sub-millimeter size are available from commercial sources (e.g.,
from Seradyn-Thermo Scientific, Indianapolis, Ind.;
Dynal-Invitrogen, Carlsbad, Calif.). Carbon nanotubes with various
functionalities can be synthesized de novo (see, e.g. Bianco et al.
in Nanomaterials for Medical Diagnosis and Therapy. pp. 85-142.
Nanotechnologies for the Live Sciences Vol. 10 Edited by Challa S.
S. R. Kumar, WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, 2007,
which is incorporated herein by reference) or may be available from
commercial sources (e.g., from Nanolab, Newton, Mass.; Swan
Chemical Inc., Lyndhurst, N.J.).
[0082] In an aspect, the device including one or more labels for
labeling target components can include an ink or dye visible with
ultraviolet, visible, near infrared, or infrared electromagnetic
energy emitted by the device. Examples of vital dyes used to stain
cells include, but are not limited to, acridin orange (stains DNA
and RNA), DiOC (3,3'-dihexyloxacarbocyanine iodide; stains
endoplasmic reticulum), rhodamine 123 (stains mitochondria), Nile
red (stains lipid vesicles), DAPI (4',6-diamidino-2-phenylindole;
stains DNA), Hoechst 33342 (stains DNA). Calcein AM and
carboxyfluorescein diacetate are examples of membrane permeable
dyes that are converted into membrane-impermeable dyes by cellular
esterases, thereby trapping them inside live cells.
[0083] The device including one or more labels configured to bind
to one or more target components can be one or more radiofrequency
identification (RFID) tags, sub-millimeter versions of which have
been described. See, e.g., Hornyak, Scientific American Magazine,
pp 68-71, February 2008, which is incorporated herein by reference.
Alternatively, the one or more label can include one or more
bokodes, millimeter sized visual tags that can be captured with a
camera. See, e.g., Mohan et al. ACM Transactions on Graphics
Proceedings of SIGGRAPH 2009, Aug. 3-7, 2009, New Orleans, which is
incorporated herein by reference.
[0084] The device including one or more labels configured to bind
one or more target components can be one or more of a radiopaque
dye or a contrast medium. Contrast agents, radiopaques, or
roentgenographic drugs used for x-ray imaging and computed
tomography (CT) include, but are not limited to, barium sulfate and
various iodine derivatives including diatrizoate meglumine,
diatrizoate sodium, iodipamide meglumine, diatrizoic acid,
ethiodized oil, iodipamide, iodixanol, iohexyl, iomeprol,
iopamidol, iopanoic acid, iophendylate, iopromide, iothalamate
meglumine, iothalamate sodium, iothalamic acid, ioversol, ioxaglate
meglumaine, or ioxaglate sodium.
[0085] The device including one or more labels configured to bind
one or more target components can include one or more contrast
agents used for magnetic resonance imaging (MRI) as exemplified by
paramagnetic and supramagnetic agents with one or more unpaired
electrons and typically including manganese, iron, or gadolinium in
their structure. Examples of MRI contrast agents containing iron
include, but are not limited to, ferumoxides (magnetite coated with
dextran), ferumoxsil (magnetite coated with siloxane), ferumoxytol,
ferumoxtran, ferucarbotran (RESOVIST), ferric chloride, ferric
ammonium citrate. Examples of MRI contrast agents containing
gadolinium include, but are not limited to, gadopentetate
dimeglumine (Gd-DTPA; MAGNEVIST), gadobutrol (GADOVIST),
gadodiamide (Gd-DTPA-BMA; OMNISCAN), gadoteridol (PROHANCE),
Gd-DOTA (DOTAREM), gadofosveset trisodium (VASOVIST),
gadoversetamide (OPTIMARK), gadobenate dimeglumine (MULTIHANCE).
Examples of MRI contrast agents containing manganese include, but
are not limited to, mangafodipir trisodium (TESLASCAN), EVP
1001-1.
Device Including Binding Components for Labels Configured to Bind
Target Components
[0086] A device is disclosed herein that can include sources of one
or more labels configured to bind to one or more target components
in the blood or lymph of a vertebrate subject. In an aspect, the
one or more labels bind directly to the one or more target
components, e.g., labels that intercalate directly and potentially
non-selectively into cellular compartments such as plasma membrane,
organelles, DNA and/or RNA. For example, cell membranes can be
readily labeled with a number of fluorescent lipophilic dyes,
examples of which include DiI
(1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine
perchlorate), DiO (3,3'-dioctadecyl-oxacarbocyanine perchlorate)
and DiA 4-(4-(dihexadecylamino)styryl)-N-methylpyridinium iodide.
In an aspect, the one or more labels can bind to a second component
that is a binding component configured to selectively bind to one
or more target components. Examples of binding components include,
but are not limited to, antibodies, antibody fragments, peptides,
oligonucleotides, aptamers, protein nucleic acids, proteins,
viruses, enzymes, receptors, bacteria, cells, cell fragments,
inorganic molecules, organic molecules, artificial binding
substrates formed by molecular imprinting or combinations
thereof.
[0087] The one or more binding components, e.g., one or more
antibodies, can be configured to bind to the label and to the
target component. Antibodies or antibody fragments for use as one
or more binding components include, but are not limited to,
monoclonal antibodies, polyclonal antibodies, Fab fragments of
monoclonal antibodies, Fab fragments of polyclonal antibodies,
Fab.sub.2 fragments of monoclonal antibodies, and Fab.sub.2
fragments of polyclonal antibodies, chimeric antibodies, non-human
antibodies, fully human antibodies, among others. Single chain or
multiple chain antigen-recognition sites can be used. Multiple
chain antigen-recognition sites can be fused or unfused. Antibodies
or fragments' thereof can be generated using standard methods. See,
e.g., Harlow & Lane, Antibodies: A Laboratory Manual, Cold
Spring Harbor Laboratory Press; 1.sup.st edition 1988, which is
incorporated herein by reference. Alternatively, an antibody or
fragment thereof directed against one or more target component can
be generated, for example, using phage display technology. See,
e.g., Kupper, et al. BMC Biotechnology 5:4, 2005, which is
incorporated herein by reference. An antibody, a fragment thereof,
or an artificial antibody, e.g., Affibody.RTM. artificial
antibodies (Affibody AB, Bromma, Sweden) can be prepared using in
silico design (Knappik et al., J. Mol. Biol. 296: 57-86, 2000,
which is incorporated herein by reference. In an aspect, antibodies
directed against one or more target components may be available
from a commercial source (from, e.g., Novus Biological, Littleton,
Colo.; Sigma-Aldrich, St. Louis, Mo.; United States Biological,
Swampscott, Mass.).
[0088] The one or more binding components, e.g., one or more
aptamers, can be configured to bind to the label and to the target
component. The aptamer can be an oligonucleotide RNA- or DNA-based
aptamer. Aptamers are artificial oligonucleotides (DNA or RNA) that
can bind to a wide variety of entities (e.g., metal ions, small
organic molecules, proteins, and cells) with high selectivity,
specificity, and affinity. Aptamers can be isolated from a large
library of 10.sup.14 to 10.sup.15 random oligonucleotide sequences
using an iterative in vitro selection procedure often termed
"systematic evolution of ligands by exponential enrichment"
(SELEX). See, e.g., Cao, et al., Current Proteomics 2:31-40, 2005;
Proske, et al., Appl. Microbiol. Biotechnol. 69:367-374, 2005;
Jayasena Clin. Chem. 45:1628-1650, 1999, each of which is
incorporated herein by reference. In general, SELEX can be used to
generate aptamers against any of a number of target components
including, but not limited to, inflammatory mediators, cancer
cells, and bacteria. See, e.g., Guthrie, et al., Methods
38:324-330, 2006; Shangguan, et al., Proc. Natl. Acad. Sci. USA.
103:11838-11843; Chen, et al., Biochem. Biophys. Res. Commun.
357:743-748, 2007, each of which is incorporated herein by
reference.
[0089] The one or more binding components, e.g., one or more
peptide-based aptamers, can be configured to bind to the label and
to the target component. Peptide aptamers are artificial proteins
in which inserted peptides are expressed as part of the primary
sequence of a structurally stable protein. See, e.g., Crawford, et
al., Brief. Funct. Genomic Proteomic 2:72-79, 2003, which is
incorporated herein by reference. Peptide aptamers can be generated
by screening a target component against yeast two-hybrid libraries,
yeast expression libraries, bacterial expression libraries and/or
retroviral libraries. Peptide aptamers can have binding affinities
comparable to antibodies.
[0090] The one or more binding components, e.g., one or more novel
peptides, can be configured to bind to the label and to the target
component. Novel peptides that bind selective targets can be
generated, for example, using phage display methodologies. See,
e.g., Spear, et al., Cancer Gene Ther. 8:506-511, 2001, which is
incorporated herein by reference. In an aspect, the phage express
novel peptides on the surface as fusion proteins in association
with a phage major or minor coat protein and can be screened for
binding interaction with one or more target components.
[0091] The one or more binding components, e.g., one or more
receptors, can be configured to bind to the label and to the target
component. All or part of a receptor can be used as a binding
component, e.g., a soluble receptor. In an aspect, the target
component can include a soluble ligand. In an aspect, the binding
component, e.g., an antibody or aptamer, configured to bind to the
label and to the target component can be configured to recognize
one or more biomolecules on the surface of the one or more target
cells. In an aspect, the binding component, e.g., an antibody or
aptamer, configured to bind to the label can be configured to
recognize one or more receptor types on the surface of target
cells. In an aspect, the target component can include a soluble
ligand. Examples of receptors include, but are not limited to,
acetylcholine receptors, adenosine receptors, adrenoceptros, GABA
receptors, angiotensin receptors, cannabinoid receptors,
cholecystokinin receptors, dopamine receptors, glucagon receptors,
glucocorticoid receptors, glutamate receptors, histamine receptors,
mineralocorticoid receptors, olfactory receptors, opioid receptors,
purinergic receptors, secretin receptors, serotonin receptors,
somatostatin receptors, steroid hormone receptors, calcium-sensing
receptor, hormone receptors, erythropoietin receptor, and
natriuretic peptide receptors. Other examples include type I
cytokine receptors such as type 1 interleukin receptors,
erythropoietin receptor, GM-CSF receptor, G-CSF receptor, growth
hormone receptor, oncostatin M receptor, leukemia inhibitory factor
receptor; type II cytokine receptors such as type II interleukin
receptors, interferon-.alpha./.beta. receptors, interferon-.gamma.
receptor; many members of the immunoglobulin superfamily such as
interleukin-1 receptor, CSF1, c-kit receptor, interleukin-18
receptor; tumor necrosis factor (TNF) receptor family such as TNF
receptor 1 (TNF-R1), TNF receptor 2 (TNF-R2), CD27, CD40, and
lymphotoxin .beta. receptor; chemokine receptors including
serpentine CCR and CXCR receptors, such as CCR1 and CXCR4, and
interleukin-8 receptor; TGF .beta. receptors such as TGF .beta.
receptor 1 and TGF .beta. receptor 2. See Ozaki and Leonard, J.
Biol. Chem. 277:29355-29358, 2002, which is incorporated herein by
reference
[0092] In some instances, the one or more binding components
configured to bind to the label and to the target component can
include one or more cellular receptors that recognize and/or bind
to bacteria. For example, the cellular receptor, CD14 which is
normally associated with monocyte/macrophages, is known to bind
lipopolysaccharide associated with gram negative bacteria as well
as lipoteichoic acid associated with the gram positive bacteria
Bacillus subtilis (see, e.g., Fan, et al. (1999) Infect. Immun. 67:
2964-2968). Other examples of cellular receptors include, but are
not limited to, adenylate cyclase (Bordatella pertussis), Gal alpha
1-4Gal-containing isoreceptors (E. coli), glycoconjugate receptors
(enteric bacteria), Lewis(b) blood group antigen receptor
(Heliobacter pylori), CR3 receptor, protein kinase receptor,
galactose N-acetylgalactosamine-inhibitable lectin receptor, and
chemokine receptor (Legionella), annexin I (Leishmania mexicana),
ActA protein (Listeria monocytogenes), meningococcal virulence
associated Opa receptors (Meningococcus), alpha5beta3 integrin
(Mycobacterium avium-M), heparin sulphate proteoglycan receptor,
CD66 receptor, integrin receptor, membrane cofactor protein, CD46,
GM1, GM2, GM3, and CD3 (Neisseria gonorrhoeae), KDEL receptor
(Pseudomonas), epidermal growth factor receptor (Samonella
typhiurium), alpha5beta1 integrin (Shigella), and nonglycosylated
J774 receptor (Streptococci) (see, e.g., U.S. Patent Application
2004/0033584 A1, which is incorporated herein by reference).
[0093] The one or more binding components, e.g., one or more
lectins, can be configured to bind to the label and to the target
component. "Lectin" has been used to define agglutinins which could
discriminate among types of red blood cells and cause
agglutination. "Lectin" can be used more generally and includes
sugar-binding proteins from many sources regardless of their
ability to agglutinate cells. Lectins have been found in plants,
viruses, microorganisms and animals. Because of the specificity
that each lectin has toward a particular carbohydrate structure,
even oligosaccharides with identical sugar compositions can be
distinguished or separated. Some lectins will bind only to
structures with mannose or glucose residues, while others can
recognize only galactose residues. Some lectins require that the
particular sugar is in a terminal non-reducing position in the
oligosaccharide, while others can bind to sugars within the
oligosaccharide chain. Some lectins do not discriminate between a
and b anomers, while others require not only the correct anomeric
structure but a specific sequence of sugars for binding. Examples
of lectins include, but are not limited to, algal lectins, e.g.,
b-prism lectin; animal lectins, e.g., tachylectin-2, C-type
lectins, C-type lectin-like, calnexin-calreticulin, capsid protein,
chitin-binding protein, ficolins, fucolectin, H-type lectins,
I-type lectins, sialoadhesin, siglec-5, siglec-7, micronemal
protein, P-type lectins, pentrxin, b-trefoil, galectins, congerins,
selenocosmia huwena lectin-I, Hcgp-39, Ym1; bacterial lectins,
e.g., Pseudomonas PA-IL, Burkholderia lectins, chromobacterium
CV-IIL, Pseudomonas PA IIL, Ralsonia RS-ILL, ADP-ribosylating
toxin, Ralstonia lectin, Clostridium hemagglutinin, botulinum
toxin, tetanus toxin, cyanobacterial lectins, FimH, GafD, PapG,
Staphylococcal enterotoxin B, toxin SSL11, toxin SSL5; fungal and
yeast lectins, e.g., Aleuria aurantia lectin, integrin-like lectin,
Agaricus lectin, Sclerotium lectin, Xerocomus lectin, Laetiporus
lectin, Marasmius oreades agglutinin, agrocybe galectin, coprinus
galectin-2, Ig-like lectins, L-type lectins; plant lectins, e.g.,
alpha-D-mannose-specific plant lectins, amaranthus antimicrobial
peptide, hevein, pokeweed lectin, Urtica dioica UD, wheat germ
WGA-1, WGA-2, WGA-3, artocarpin, artocarpus hirsute AHL, banana
lectin, Calsepa, heltuba, jacalin, Maclura pomifera MPA, MornigaM,
Parkia lectins, abrin-a, abrus agglutinin, amaranthin, castor bean
ricin B, ebulin, mistletoe lectin, TKL-1, cyanovirin-N homolog, and
various legume lectins; and viral lectins, e.g., capsid protein,
coat protein, fiber knob, hemagglutinin, and tailspike protein.
See, e.g., E. Bettler, R. Loris, A. Imberty "3D-Lectin database: A
web site for images and structural information on lectins" 3rd
Electronic Glycoscience Conference, The interne and World Wide Web,
6-17 Oct. 1997; http://www.cermav.cnrs.fr/lectines/, which is
incorporated herein by reference.
[0094] The one or more binding components configured to bind to the
label can include one or more artificial binding substrates formed
by the process of molecular imprinting. In the process of molecular
imprinting, a template, e.g., target component, is combined with
functional monomers which upon cross-linking form a polymer matrix
that surrounds the template. See, e.g., Alexander, et al., J. Mol.
Recognit. 19:106-180, 2006, which is incorporated herein by
reference. Removal of the template leaves a stable cavity in the
polymer matrix that is complementary in size and shape to the
template. In an aspect, functional monomers of acrylamide and
ethylene glycol dimethacrylate can be mixed with one or more target
components in the presence of a photoinitiator and ultraviolet
irradiation used to cross-link the monomers. The resulting polymer
can be crushed or ground into smaller pieces and washed to remove
the one or more target components, leaving a particulate matrix
material capable of binding one or more target components. Examples
of other functional monomers, cross-linkers and initiators can be
used to generate an artificial binding substrate are provided. See,
e.g., U.S. Pat. No. 7,319,038; Alexander, et al., J. Mol. Recognit.
19:106-180, 2006, each of which is incorporated herein by
reference. In an aspect, hydrogels can be used for molecular
imprinting. See, e.g., Byrne et al., "Molecular imprinting within
hydrogels", Advanced Drug Delivery Reviews, 54: 149-161, 2002,
which is incorporated herein by reference. Other examples of
synthetic binders are provided. See, e.g., U.S. Pat. Nos.
6,255,461; 5,804,563; 6,797,522; 6,670,427; and 5,831,012; and U.S.
Patent Application 20040018508; and Ye and Haupt, Anal Bioanal
Chem. 378: 1887-1897, 2004; Peppas and Huang, Pharm Res. 19:
578-587 2002, each of which is incorporated herein by
reference.
[0095] In an aspect, the binding component configured to bind to
the label and to the target component can be configured to
recognize other biomolecules on the surface of target cells
including, but not limited to, various CD (cluster of
designation/cluster of differentiation) markers, intergrins, ion
channels, ATPases, cell adhesion molecules, integral membrane
glycoproteins, immunoglobulins, transporters. The one or more
binding components can be configured to recognize components of
cell surface biomolecules including amino acid sequence and
oligosaccharide modifications.
[0096] In an aspect, the binding component configured to bind to
the label and to the target component can be configured to
recognize a biomolecule associated with a tumor cell. Examples of
tumor associated components can include, but are not limited to,
BLyS receptor, carcinoembryonic antigen (CA-125), CD25, CD34, CD33
and CD123 (acute myeloid leukemia), CD20 (chronic lymphocytic
leukemia), CD19 and CD22 (acute lymphoblastic leukemia), CD30,
CD40, CD70, CD133, 57 kD cytokeratin, epithelial specific antigen,
epithelial cell adhesion molecule (EpCAM), extracellular matrix
glycoprotein tenascin, Fas/CD95, folate receptor, gastrin-releasing
peptide-like receptors, hepatocyte specific antigen, human gastric
mucin, human milk fat globule, lymphatic endothelial cell marker,
matrix metalloproteinase 9, melan A, melanoma marker, mesothelin,
mucin glycoproteins (e.g., MUC1, MUC2, MUC4, MUC5AC, MUC6),
prostate specific antigen, prostatic acid phosphatase, PTEN, renal
cell carcinoma marker, RGD-peptide binding integrins, sialyl Lewis
A, six-transmembrane epithelial antigen of the prostate (STEAP),
TNF receptor, TRAIL receptor, tyrosinase, villin. Other tumor
associated antigens include, but are not limited to, alpha
fetoprotein, apolipoprotein D, clusterin, chromogranin A,
myeloperoxidase, MyoD1 myoglobin placental alkaline phosphatase
c-fos, homeobox genes.
[0097] In an aspect, the binding component configured to bind to
the label and to the target component can be configured to
recognize a biomolecule associated with the surface of a pathogen,
e.g., bacteria, a virus, a fungus, or a parasite. The biomolecule
can be one or more components of the bacterial outer membrane, cell
wall, and/or cytoplasmic membrane. Examples of components
associated with the bacterial outer membrane of Gram-negative
bacteria include, but are not limited to, lipopolysaccaride and OMP
(outer membrane protein) porins, the latter of which are
exemplified by OmpC, OmpF and PhoP of E. coli. Examples of
components associated with the bacterial cell wall of both
Gram-positive and Gram-negative bacterial include, but are not
limited to, peptidoglycans polymers composed of an alternating
sequence of N-acetylglucoamine and N-acetyl-muraminic acid and
crosslinked by amino acids and amino acid derivatives. Examples of
components associated with the bacterial cytoplasmic membrane
include, but are not limited to, the MPA1-C (also called
polysaccharide copolymerase, PCP2a) family of proteins, the MPA2
family of proteins, and the ABC bacteriocin exporter accessory
protein (BEA) family of proteins. Other examples of components
associated with bacteria include, but are not limited to,
transporters, e.g., sugar porter (major facilitator superfamily),
amino-acid/polyamine/organocation (APC) superfamily, cation
diffusion facilitator, resistance-nodulation-division type
transporter, SecDF, calcium:cation antiporter, inorganic phosphate
transporter, monovalent cation:proton antiporter-1, monovalent
cation:proton antiporter-2, potassium transporter,
nucleobase:cation symporter-2, formate-nitrite transporter,
divalent anion:sodium symporter, ammonium transporter, and
multi-antimicrobial extrusion; channels, e.g., major intrinsic
protein, chloride channel, and metal ion transporter; and primary
active transporters, e.g., P-type ATPase, arsenite-antimonite
efflux, Type II secretory pathway (SecY), and sodium-transporting
carboxylic acid decarboxylase. A number of other potential
components associated with bacteria have been described in Chung,
et al., J. Bacteriology 183:1012-1021, 2001, which is incorporated
herein by reference.
[0098] In an aspect, the binding component configured to bind to
the label and to the target component can be configured to
recognize a biomolecule associated with a blood cell infected with
a pathogen. For example, red blood cells infected with P.
falciparum can be distinguished from normal red blood cells by
changes in surface protein expression including expression on the
red blood cell surface of the parasite derived protein P.
falciparum erythrocyte membrane protein (PfEMP1). See, e.g.,
Horata, et al., Malaria J. 8:184, 2009, which is incorporated
herein by reference.
Device Including Labels Linked to Binding Components to Elicit a
Signal Response
[0099] The device can include at least one first reservoir that
includes one or more labels that bind one or more target components
from the blood or lymph of the vertebrate subject to elicit a
signal response from the labeled binding component. The one or more
labels can include a binding component configured to bind
selectively to one or more target components. The one or more
labels that bind to one or more target components can be linked to
one or more binding components using a variety of methods
including, but not limited to, one or more of a chemical
cross-link, a streptavidin/biotin interaction (Thermo Fisher
Scientific, Rockford, Ill.), his-tagged protein on nickel-NTA resin
affinity column (Thermo Fisher Scientific, Rockford, Ill.), a
fusion protein construct, a common substrate, or a combination
thereof. In an aspect, the label and the binding component are
directly associated with one another through chemical
cross-linking, non-covalent linking, or synthesis as a single
molecule. In another aspect, the label and the binding component
are indirectly associated. In this instance, the label and the
binding component are separately attached to a mobile substrate
such as, for example, a bead or other particle-like substrate
capable of being released into the blood or lymph of the vertebrate
subject. The particle-like substrate can include a bead, a vesicle,
a cell, a carbon nanotube, or other similar structure.
[0100] In an aspect, the label can be conjugated to the binding
component using one or more of a cross linking agent. In general,
any of a number of cross linking agents can be used to conjugate an
appropriately derivatized label to an appropriately derivatized or
functionalized binding component. For example, a fluorescent dye,
e.g., rhodamine, derivatized with succinimidyl ester (from, e.g.,
Invitrogen, Carlsbad, Calif.) will react efficiently with primary
amines of proteins, e.g., antibodies, to generate a stable
fluorescent dye-protein conjugate. See, e.g., Hama et al., Cancer
Res. 67:2791-2799, 2007, which is incorporated herein by reference.
As another example, an antibody for use in a label configured to
bind to a target component can be conjugated with one or more
quantum dots via an amine-thiol linkage using amine-derivatized,
poly-ethylene glycol coated quantum dots and the amine-thiol
crosslinker SMCC using a commercially available kit (Qdot.RTM.
Antibody Conjugation Kit, Invitrogen, Carlsbad, Calif.). Similarly,
various methods are available for attaching quantum dots to a DNA
backbone of an aptamer such as, for example, covalent linkage of
amine-modified DNA to carboxylated quantum dots. For example,
carboxy quantum dots (from, e.g., Quantum Dot Corporation, Hayward,
Calif., USA) can be attached to an aptamer through a C6 amino
modifier placed on either the 5 prime or 3 prime end of the aptamer
sequence. Magnetic beads derivatized with carboxylic acid, amine
groups or tosylactivated for cross-linking to proteins and
appropriately derivatized oligonucleotides are also commercially
available (from, e.g., Dynal Biotech, Brown Deer, Wis.). Quantum
dots, fluorescent dyes, and magnetic particles derivatized for
cross-linking to antibodies, aptamers or other biomolecules are
available from a number of commercial sources (from, e.g.,
Invitrogen, Carlsbad, Calif.; Seradyn-Thermo Scientific,
Indianapolis, Ind.; Sigma-Aldrich, St. Louis, Mo.).
[0101] In an aspect, the derivatized fluorescent dye itself can be
used to label target cells. For example, cells can be stably
labeled with 5,6-carboxyfluorescein diacetate succinimidyl ester, a
derivatized fluorescent dye capable of entering a cell and once
inside a cell, binding to primary amines associated with
cytoplasmic proteins. See, e.g., Humphreys et al, Toxicol. Sci.
73:80-89, 2003, which is incorporated herein by reference.
[0102] Examples of cross linking agents for cross linking one or
more of a label with one or more of a binding component include,
but are not limited to, primary amine/primary amine linkers such as
DMS (dimethyl suberimidate), DMP (dimethyl pimelimidate), DSS
(disuccinimidyl suberate), DST (disuccinimidyl tartate), Sulfo DST
(sulfodisuccinimidyl tartate), DSP (dithiobis(succinimidyl
propionate) and sulfhydryl/sulfhydryl linkers such as DPDPB
(1,4-di-(3'-[2'pyridyldithio]-propionamido) butane); primary
amine/sulfhydryl linkers such as MBS
(m-maleimidobenzoyl-N-hydroxysuccinimide ester), Sulfo MBS
(m-maleimidobenzoyl-N-hydroxysulfosuccinimide), Sulfo GMBS
(N-.gamma.-maleimidobutyryloxysulfosuccinimide ester), EMCS
(N-(epsilon-maleimidocaproyloxy)succinimide ester), Sulfo EMCS
(N-(epsilon-maleimidocaproyloxy)sulfo succinimide), SMCC
(succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate), SMPB
(succinimidyl 4-(rho-maleimidophenyl)butyrate), Sulfo SIAB
(N-sulfosuccinimidyl(4-iodoacetyl)aminobenzoate),
cyclohexane-1-carboxylate), and MAL-PEG-NHS (maleimide PEG
N-hydroxysuccinimide ester); sulfhydryl/hydroxyl linkers such as
PMPI (N-rho-maleimidophenyl)isocyanate; sulfhydryl/carbohydrate
linkers such as EMCH (N-(epsilon-maleimidocaproic acid)hydrazide);
and amine/carboxyl linkers such as EDC
(1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride).
[0103] In an aspect, the label can be conjugated to the binding
component using one or more interactions between biotin and avidin,
streptavidin or derivatives thereof. Streptavidin and avidin are
multivalent proteins capable of binding multiple biotin subunits
with high affinity and as such can be used as linking molecules
between one or more biotinylated label and one or more biotinylated
binding component. For example, a biotinylated fluorescent label,
e.g., biotin-4-fluorescein (from, e.g., Invitrogen, Carlsbad,
Calif.) can be linked to a biotinylated antibody through a
streptavidin bridge. An antibody or other protein-based binding
component can be biotinylated using an amine reactive biotinylation
reagent such as, for example, EZ-Link Sulfo-NHS-SS-Biotin
(sulfosuccinimidyl 2-(biotinamido)-ethyl-1,3-dithiopropionate;
Pierce-Thermo Scientific, Rockford, Ill., USA; see, e.g., Jaiswal,
et al. Nature Biotech. 21:47-51, 2003, which is incorporated herein
by reference). Similarly, a biotinylated label can be linked to a
biotinylated oligonucleotide aptamer through a streptavidin bridge.
An aptamer or other nucleotide-based binding component can be
biotinylated by introducing a biotinylated nucleotide, e.g.,
biotin-5-deoxycytidine-5-triphosphate (from, e.g., ChemCyte, Inc.,
San Diego, Calif.) into the aptamer sequence during in vitro
transcription. The biotinylated label is reacted with the
biotinylated binding component in the presence of streptavidin to
generate a label for binding to a target component. Radioactive
labels, e.g., iodine-125, can also be biotinylated. See, e.g.,
Garlick & Giese. J. Biol. Chem. 263:210-215, 1988, which is
incorporated herein by reference.
[0104] Alternatively, the label or the binding component can be
modified with streptavidin, avidin, or derivative thereof and
directly bound to a biotinylated label or binding component. In an
aspect, the label is modified with streptavidin and combined with a
biotinylated binding component. For example, streptavidin modified
quantum dots (available from, e.g., Quantum Dot Corporation,
Hayward, Calif., USA) can be attached to an aptamer through a
biotin modification to the 5-prime end of the aptamer sequence.
See, e.g., Cady et al. Mol. Cell. Probes 21:116-124, 2007, which is
incorporated herein by reference. Examples of other streptavidin
modified fluorescent dyes are available (from, e.g., PerkinElmer,
Waltham, Mass.; Alpha Diagnostic Intl. Inc., San Antonio, Tex.).
Streptavidin modified magnetic beads are also commercially
available (e.g., Dynabeads.RTM. MyOne.TM. Streptavidin, Dynal
Biotech, Brown Deer, Wis.). In another aspect, the binding
component can contain all or part of the streptavidin protein for
use in binding to a biotin modified label. For example, cDNA
sequence encoding all or part of an antibody or other
protein/peptide can be genetically modified to contain all or part
of the streptavidin gene using standard cloning procedures,
resulting in a streptavidin-antibody fusion protein. See, e.g.,
Koo, et al. Appl. Environ. Microbiol. 64:2497-2502, 1998, which is
incorporated herein by reference. The streptavidin modified binding
component can subsequently be combined with one or more of a
biotinylated label to generate label configured to bind one or more
target components.
[0105] In an aspect, the label, e.g., radioactive label or
fluorescent label, can be incorporated into the label at the time
of synthesis. For example, radiolabeled nucleotides or radiolabeled
amino acids can be incorporated respectively into an
oligonucleotide aptamer or protein antibody or other protein-based
binding component during synthesis using standard methods.
Alternatively, the label can be a fusion protein with a binding
component, e.g., antibody, peptide ligand, or receptor, designed to
bind to or associate with a target component and a label component
including all or part of green fluorescent protein (GFP) derived
from Aequorea victoria jellyfish or yellow, red and blue
fluorescing derivatives thereof. A number of expression constructs
for generating recombinant GFP fusion proteins are available from
commercial sources (from, e.g., Invitrogen, Carlsbad, Calif.).
[0106] In another aspect, the label configured to bind to one or
more target components can be a DNA construct encoding a
fluorescent protein and inserted into one or more target cells in
the blood or lymph of a vertebrate subject. For example,
baculovirus expression constructs are available which when
introduced into a cell induce expression of specific fluorescent
proteins, e.g., Cellular Lights.TM. fluorescent proteins and
Organelle Lights.TM. fluorescent proteins (from Invitrogen,
Carlsbad, Calif.).
[0107] Methods have also been described for incorporating the label
configured to bind to one or more target components wherein gold
nanoparticles can be linked to aptamers for use in reflectance
imaging applications, in which the aptamer is derivatized with a
5-prime thiol group and interacted with colloidal gold
nanoparticles (from, e.g., Ted Pella, Inc., Redding, Calif.). See,
e.g., Javier et al. Bioconjugate Chem. 19:1309-1312, 2008, which is
incorporated herein by reference.
[0108] In an aspect, the detectable label including a binding
component may be available from a commercial source. For example,
lectins concanavalin A and wheat germ agglutinin are available
conjugated to Alexa fluors, Marina Blue, AMCA, Oregon Green,
tetramethylrhodamine, Texas Red, fluorescein (from, Invitrogen,
Carlsbad, Calif.). Other lectins conjugated to fluorescent dyes are
available including Phaseolus vulgaris lectin (PHA-L), Arachis
hypogaea lectin (PNA), Helix pomatia agglutinin (HPA), Soybean
agglutinin (SBA), and lectins from Griffonia simplicifolia (from,
Invitrogen, Carlsbad, Calif.). Magnetic beads with an antibody to
the human epithelial antigen, EpCAM (epithelial cell adhesion
molecule) are commercially available (from, e.g., Dynal Biotech,
Brown Deer, Wis.). EpCAM can be used to selectively bind
circulating tumor cells of epithelial origin in the blood or lymph
of a vertebrate subject. Anti-CA-125 (anti-carcinoembryonic
antigen) antibodies can be used to selectively bind circulating
tumor cells of ovarian cancer origin in the blood or lymph of a
mammalian subject. Anti-CA125 antibodies can be conjugated to
rhodamine-X (Invitrogen, Eugene, Oreg.). Anti-FR (anti-folate
receptor) antibodies and folate-FITC, folate-Tc99m can be used to
selectively bind circulating tumor cells that overexpress folate
receptors, e.g., ovarian cancer cells, and circulating tumor cells
in the blood or lymph of a mammalian subject. Endocyte, Inc., West
Lafayette, Ind. See, e.g., He, et al., Proc. Natl. Acad. Sci. USA
104: 11760-11765, 2007, which is incorporated herein by
reference.
Device Including Activatable labels for Labeling Target
Components
[0109] The device can include at least one first reservoir that
includes one or more labels that bind one or more target components
from the blood or lymph of the vertebrate subject. The device can
include one or more energy sources configured to provide energy to
elicit one or more signal responses associated with one or more
labeled target components. The one or more labels that bind to one
or more target components can be one or more activatable labels
configured to controllably emit a measurable signal in response to
binding a target component. In an aspect, the one or more
activatable labels can emit a measurable signal only when a target
component is bound. Alternatively, the one or more activatable
labels can emit a first measurable signal in the absence of a bound
target component and a second measurable signal in the presence of
a bound target component.
[0110] In an aspect, the activatable label can be configured such
that binding of the target component to the binding component of
the label results in a conformational change in the label that can
be measured using fluorescence resonance energy transfer (FRET).
FRET is a distance-dependent interaction between the electronic
excited states of two dye molecules in which excitation is
transferred from a donor molecule to an acceptor molecule without
emission of a photon. In an aspect, interaction of a donor molecule
with an acceptor molecule can lead to a shift in the emission
wavelength associated with excitation of the acceptor molecule. In
an aspect, interaction of a donor molecule with an acceptor
molecule can lead to quenching of the donor emission. The binding
component of the activatable label configured to bind one or more
target components can include at least one donor molecule and at
least one acceptor molecule. In this configuration, binding of one
or more target components to the binding component of the
activatable label results in a conformational change in the binding
component and results in a change in the distance between the donor
and acceptor molecules and a change in measurable label, e.g.,
fluorescence.
[0111] A variety of donor and acceptor fluorophore pairs can be
considered for FRET including, but not limited to, fluorescein and
tetramethylrhodamine; IAEDANS and fluorescein; fluorescein and
fluorescein; and BODIPY FL and BODIPY FL. A number of Alexa Fluor
(AF) fluorophores (Molecular Probes-Invitrogen, Carlsbad, Calif.,
USA) can be paired with other AF fluorophores for use in FRET. Some
examples include, but are not limited, to AF 350 with AF 488; AF
488 with AF 546, AF 555, AF 568, or AF 647; AF 546 with AF 568, AF
594, or AF 647; AF 555 with AF594 or AF647; AF 568 with AF6456; and
AF594 with AF 647.
[0112] The cyanine dyes Cy3, Cy5, Cy5.5 and Cy7, which emit in the
red and far red wavelength range (>550 nm), offer a number of
advantages for FRET-based detection systems. Their emission range
is such that background fluorescence is often reduced and
relatively large distances (>100 .ANG.) can be measured as a
result of the high extinction coefficients and good quantum yields.
For example, Cy3, which emits maximally at 570 nm and Cy5, which
emits at 670 nm, can be used as a donor-acceptor pair. When the Cy3
and Cy5 are not proximal to one another, excitation at 540 nm
results only in the emission of light by Cy3 at 590 nm. In
contrast, when Cy3 and Cy5 are brought into proximity by a
conformation change in an aptamer, antibody, or receptor, for
example, excitation at 540 nm results in an emission at 680 nm.
[0113] Quenching dyes can be used as part of the activatable label
to quench the fluorescence of visible light-excited fluorophores.
Examples of quenching dyes include, but are not limited, to DABCYL,
the non-fluorescing diarylrhodamine derivative dyes QSY 7, QSY 9
and QSY 21 (Molecular Probes, Carlsbad, Calif., USA), the
non-fluorescing Black Hole Quenchers BHQ0, BHQ1, BHQ2, and BHQ3
(Biosearch Technologies, Inc., Novato, Calif., USA) and Eclipse
(Applera Corp., Norwalk, Conn., USA). A variety of donor
fluorophore and quencher pairs can be considered for FRET
associated with the label including, but not limited to,
fluorescein with DABCYL; EDANS with DABCYL; or fluorescein with QSY
7 and QSY 9. In general, QSY 7 and QSY 9 dyes efficiently quench
the fluorescence emission of donor dyes including blue-fluorescent
coumarins, green- or orange-fluorescent dyes, and conjugates of the
Texas Red and Alexa Fluor 594 dyes. QSY 21 dye efficiently quenches
all red-fluorescent dyes. A number of the Alexa Fluor (AF)
fluorophores (Molecular Probes-Invitrogen, Carlsbad, Calif., USA)
can be paired with quenching molecules as follows: AF 350 with QSY
35 or DABCYL; AF 488 with QSY 35, DABCYL, QSY7 or QSY9; AF 546 with
QSY 35, DABCYL, QSY7 or QSY9; AF 555 with QSY7 or QSY9; AF 568 with
QSY7, QSY9 or QSY21; AF 594 with QSY21; and AF 647 with QSY 21.
[0114] The activatable label can include a binding component that
is an RNA or DNA oligonucleotide-based aptamer with one or more
fluorescent tags and one or more quenching tags. Upon binding of a
target component, the aptamer undergoes a conformational shift such
that the distance between the donor fluorophore and the acceptor
fluorophore or quencher is altered, leading to a change in
measurable fluorescence. Aptamers can be isolated from a large
library of 10.sup.14 to 10.sup.15 random oligonucleotide sequences
using an iterative in vitro selection procedure often termed
"systematic evolution of ligands by exponential enrichment"
(SELEX). Aptamers can be generated against virtually any class of
molecules including cells (e.g., cancer cells, bacteria, and
parasites), proteins (e.g., hormones), and chemicals (e.g.,
codeine, cocaine). See, e.g., Shangguan, et al., Proc. Natl. Acad.
Sci. USA. 103:11838-11843; Chen, et al., Biochem. Biophys. Res.
Commun. 357:743-748, 2007; Ulrich, et al., J. Biol. Chem.
277:20756-20762, 2002; Cao, et al. Current Proteomics 2:31-40,
2005; Proske, et al. Appl. Microbiol. Biotechnol. 69:367-374, 2005,
Win, et al. Nucleic Acids Res. 34:5670-5682, 2006, each of which is
incorporated herein by reference. For example, an aptamer that
selectively binds cocaine can be generated using SELEX as described
above and modified to incorporate a fluorophore such as, for
example, fluorescein and a quencher such as, for example,
4-[4'-((dimethylamino)phenyl)azo-]benzoic acid (DABCYL). See, e.g.,
Strojanovic, et al. J. Am. Chem. Soc. 123:4928-4931, 2001, which is
incorporated herein by reference. In this instance, binding of
cocaine to the aptamer induces a conformational change in the
aptamer that causes the fluorophore and the quencher to move closer
in proximity. As such, the presence of cocaine can be measured as a
function of the decrease in or quenching of the fluorescein
emission at a wavelength of 518 nm.
[0115] Semiconductor quantum dots (QDs) with various
excitation/emission wavelength properties can be used to label an
aptamer-based binding component. Various methods are available for
attaching quantum dots to the DNA backbone of an aptamer such as,
for example, covalent linkage of amine-modified DNA to carboxylated
quantum dots and linkage of biotinylated DNA to streptavidin
modified quantum dots. See, e.g., Cady, et al. J. Mol. Cell. Probes
21:116-124, 2007, which is incorporated herein by reference. For
example, carboxy quantum dots (from, e.g., Quantum Dot Corporation,
Hayward, Calif., USA) can be attached to an aptamer through a C6
amino modifier placed on either the 5 prime or 3 prime end of the
aptamer sequence. Alternatively, streptavidin quantum dots (from,
e.g., Quantum Dot Corporation, Hayward, Calif., USA) can be
attached to an aptamer through a biotin attached to the 5-prime end
of the aptamer sequence.
[0116] The fluorophores can be attached to various chemical
moieties that allow for attachment at various sites within the
aptamer. For example, 3'-DABCYL CPG can be used to place the
fluorophore DABCYL at the 3 prime terminus of the aptamer whereas
5'-DABCYL phosphoramidite can be used to place DABSYL at the 5
prime terminus of the aptamer (see, e.g., product information at
Glen Research, Sterling, Va.). DABCYL deoxythymidine (dT) can be
used to place DABCYL within the body of the aptamer sequence.
Labeling aptamers with appropriate commercially available
fluorophores can be achieved following instructions provided by the
respective manufacturer. Alternatively, custom made aptamer-based
molecular beacons are available from commercial sources (from,
e.g., Biosearch Technologies, Inc., Novato, Calif., USA).
[0117] In an aspect, an aptamer can have a label in a region of the
molecule known to undergo conformational change upon binding a
target component that leads to an increase in fluorescence
intensity. An aptamer of this sort can be selected using an in
vitro selection process with fluorescently labeled aptamers. See,
e.g., Jhaveri, et al. Nature Biotech. 18:1293-1297, 2000, which is
incorporated herein by reference. A pool of RNA molecules is
generated in which the random sequence region (45-60 residues) is
skewed such that one of the residues, uridine, for example, is
disproportionately underrepresented. The three to four randomly
placed uridine residues are substituted with fluorescein-12-UTP,
Cascade Blue-7-UTP, Texas Red-5-UTP, and/or Rhodamine Green-5-UTP
during in vitro transcription. The labeled pool of RNA molecules
are screened against a target component by passing the labeled pool
over a column matrix or other matrix to which the target component
is attached. Those RNA molecules that bind with high affinity to
the target component are screened for their fluorescence signaling
properties in response to binding of the target component. For
example, the baseline fluorescence intensity is measured for RNA
aptamer molecules labeled with fluorescein-12-UTP (excitation
maxima 494 nm, emission maxima 521 nm) or Rhodamine Green-5-UTP
(excitation maxima 505 nm, emission maxima 533 nm), for example,
then re-measured in response to increasing concentrations of the
target component. As such, fluorescent aptamers can be selected
that exhibit a 100-200% increase in fluorescence intensity in
response to target binding. See, e.g., Jhaveri, et al. Nature
Biotech. 18:1293-1297, 2000, which is incorporated herein by
reference.
[0118] In an aspect, the label configured to bind to one or more
target components can include a binding component that is an
antibody with one or more donor fluorophore and one or more
acceptor fluorophore or quencher. Upon binding of a target
component, the antibody undergoes a conformational shift such that
the distance between the donor fluorophore and the acceptor
fluorophore or quencher is altered, leading to a change in
measurable signal, i.e., fluorescence. The antibody can be designed
to emit a shift in emission wavelength, for example, in response to
binding a target component. An antibody exhibiting a shift in
fluorescent signal in response to binding of a target component,
i.e., antigen, can be generated by labeling the antibody with a
solvent-sensitive fluorophore, e.g., dansyl chloride
(5-dimethylaminonaphthalene-1-sulfonyl chloride). See, e.g.,
Brennan J. Fluor. 9:295-312, 1999, which is incorporated herein by
reference. The antibody is labeled such that binding of the target
component to the antibody shields the solvent sensitive fluorescent
label near the active binding site from the solvent water,
resulting in a 3-5 fold increase in fluorescence intensity. See,
e.g., Bright, et al. Anal. Chem. 62:1065-1069, 1990, which is
incorporated herein by reference.
[0119] In an aspect, the label configured to bind to one or more
target components can include a binding component that is an
antibody that signals binding of a target component using FRET and
a flexible arm. For example, an antibody can include a donor
fluorophore near the binding site of the target component as well
as a flexible arm containing an analog of the target component that
is labeled with a quencher and recognized by the antibody. See,
e.g. U.S. Patent Application 2006/0172318 A1, which is incorporated
herein by reference. As the labeled target component analog moves
into proximity to the labeled active site, a baseline FRET signal
can be measured. A measurable change in the FRET signal is detected
when the analog is competitively displaced by the actual target
component. The flexible arm can be composed of DNA, RNA, polymers,
protein nucleic acid (PNA), peptides, protein or oligosaccharide.
For example, an amino functionalized DNA arm can be treated with a
bifunctional NHS-ester activated Cy3.5 dye to add a fluorescent tag
to the flexible arm. The analog of the target component is modified
with a monoamine and interacted with the bifunctional NHS-ester and
attached to the DNA flexible arm. See, e.g., U.S. Patent
Application 2006/0172318 A1, which is incorporated herein by
reference. The flexible arm can be attached directly to the
antibody through a thiol-maleimide linkage such that the DNA
flexible arm is modified with a thiol group at one end and linked
via maleimide to one or more cysteine groups on the antibody.
Alternatively, the flexible arm can be attached to a protein, for
example, that is adjacent to the antibody or to which the antibody
is bound.
Device Including First Reservoir for Storing and Releasing
Label
[0120] The device can include at least one first reservoir that
includes one or more labels that bind one or more target components
from the blood or lymph of the vertebrate subject. The device can
include at least one first reservoir that stores and controllably
releases the one or more labels configured to bind to the one or
more target components. In an aspect, the device can include a
single first reservoir that stores one or more labels configured to
bind to one or more target components. In another aspect, the
device can include multiple first reservoirs wherein each multiple
first reservoir can store one or more labels that bind to one or
more target components. The at least one first reservoir can be an
integral part of the device. Alternatively, the at least one first
reservoir can be a separate part of the device, located proximal to
or at a distance from the main body of the device, but in wired or
wireless communication with the main body of the device.
[0121] The device can include one or more labels configured to bind
to one or more target component so that the one or more labels can
be released from the at least one first reservoir by one or more
release mechanisms. In an aspect, the one or more labels can be
continuously released from at least one first reservoir at a
constant rate over time. For example, the release mechanism can use
one or more of a slow release, controlled release, or extended
release biodegradable composition that dissolves or breaks down
over time. Examples of slow release, controlled release, or
extended release compositions include, but are not limited to,
hydrogels, polymers, gelled and/or cross-linked water swellable
polyolefins, polycarbonates, polyesters, polyamides, polyethers,
polyepoxides and polyurethanes such as, for example,
poly(acrylamide), poly(2-hydroxyethyl acrylate),
poly(2-hydroxypropyl acrylate), poly(N-vinyl-2-pyrrolidone),
poly(n-methylol acrylamide), poly(diacetone acrylamide),
poly(2-hydroxylethyl methacrylate), poly(allyl alcohol). Other
suitable polymers include, but are not limited to, cellulose
ethers, methyl cellulose ethers, cellulose and hydroxylated
cellulose, methyl cellulose and hydroxylated methyl cellulose, gums
such as guar, locust, karaya, xanthan gelatin, and derivatives
thereof. The rate of dissolution of the composition containing the
labels can be monitored using an impedance-based sensor. See, e.g.,
Johnson et al., in J. Electrochem Soc. 152:H6-H11, 2005, which is
incorporated herein by reference.
[0122] In another aspect, the device can include the one or more
labels configured to bind one or more target components, wherein
the one or more labels can be controllably released from the at
least one first reservoir and can include periods of release
followed by periods of non-release. Controlled release from at
least one first reservoir can include release mechanisms that
reversibly open and close a portion of the first reservoir. The
release mechanism can include a variety of different types of
release mechanisms, including, for example, a controllable valve.
Various examples of micro valves or microelectromechanical systems
(MEMS) valves for controlling fluid flow have been described. See,
e.g., Luckevich M. Valve World, May 2007, pp. 79-83; Givrad T K.,
et al., Proceedings of BIOMed 2008, 3.sup.rd Frontiers in
Biomedical Devices Conference. Jun. 18-20, 2008, Irvine, Calif.,
USA; U.S. Pat. Nos. 6,612,535; 7,124,773, each of which is
incorporated herein by reference.
[0123] In an aspect, the device can include the at least one first
reservoir covered with a material that can be controllably removed
or punctured to release one or more labels. The cover material can
be responsive to a directly applied stimulus (e.g., an applied
voltage or potential) or to a change in the local environment of
the device (e.g., local pH change), or any of a number of other
stimuli including, but not limited to, heat, light (e.g., laser),
and magnetic field. See, e.g., U.S. Pat. No. 6,808,522; Grayson, R.
et al., Proceedings of IEEE 92:6-21, 2004, each of which is
incorporated herein by reference. As an example, the at least one
first reservoir can be an array of microreservoirs on a microchip
in which each aliquot of one or more labels are contained in its
own reservoir and capped by an environmentally sensitive material.
In an aspect, the microreservoirs can be capped with a gold
membrane which is weakened and ruptured by electrochemical
dissolution in response to application of an anode voltage to the
membrane in the presence of chloride ions, resulting in release of
contents of the microreservoir as described in U.S. Pat. No.
5,797,898 and in Prescott, et al., Nat. Biotech., 24:437-438, 2006,
each of which is incorporated herein by reference. Alternatively,
the microreservoirs can be capped by a temperature sensitive
material which can be ruptured in response to selective application
of heat to one or more of the reservoirs as described in U.S. Pat.
No. 6,669,683, which is incorporated herein by reference. Wireless
induction of a voltage or thermal trigger, for example, to a given
reservoir of a microarray of reservoirs by a vertebrate subject
would enable on-demand release of one or more labels or reactive
components from the reservoirs.
[0124] In some instances, the device can include the at least one
first reservoir wherein the at least one first reservoir can
incorporate a natural and/or synthetic stimulus-responsive hydrogel
or polymer which changes confirmation rapidly and reversibly in
response to environmental stimuli, for example, temperature, pH,
ionic strength, electrical potential, light, magnetic field or
ultrasound. See, e.g., Stubbe, et al., Pharmaceutical Res.,
21:1732-1740, 2004, which is incorporated herein by reference.
Examples of polymers are described in U.S. Pat. Nos. 5,830,207;
6,720,402; and 7,033,571, each of which is incorporated herein by
reference. In some instances, the one or more labels can be
dissolved or dispersed in the hydrogel or polymer. Alternatively, a
hydrogel and/or other stimulus-responsive polymer can be
incorporated into the release mechanism. For example, a hydrogel or
other polymer or other smart material can be used as an
environmentally sensitive actuator to control flow of an agent out
of an implantable device as described in U.S. Pat. Nos. 6,416,495;
6,571,125; and 6,755,621, each of which is incorporated herein by
reference. As such, the at least one first reservoir can
incorporate a hydrogel or other polymer that modulates delivery of
a one or more labels in response to a trigger from the
sensor-informed controller.
[0125] In some instances, the one or more labels configured to bind
one or more target components can be released from the at least one
first reservoir as a predetermined dosage. For example, the one or
more labels can be administered using continuous infusion.
Alternatively, the release of one or more labels from the at least
one first reservoir can be linked to a timing mechanism associated
with the device. For example, the timing mechanism can instruct
release of one or more labels at a given time of day, a given time
of week, a given time of month, and/or a given time of year. The
timing mechanism can be linked to input from the one or more
sensors, releasing one or more labels at one or more predetermined
time following detection of one or more target components.
[0126] The release of one or more labels from the first reservoir
can be programmable, having on/off and/or variable delivery rates
based on either external or internal control. External control can
be mediated by manual manipulation of a hand-operated pulsative
pump with one-way valves associated with a device implanted near
the surface of the skin, for example. Alternatively, external
control can be mediated by remote control through an
electromagnetic wireless signal such as, for example, infrared or
radio waves that are able to trigger an electrical stimulus within
the implanted device. Examples of remote control drug delivery
devices are described in U.S. Pat. Nos. 5,928,195; 6,454,759; and
6,551,235, each of which is incorporated herein by reference. As
such, one or more label can be delivered by continuous infusion in
response to an "on" trigger and stopped in response to an "off"
trigger, for example. Alternatively, one or more label can be
delivered as a microbolus, for example, in response to an "on"
trigger as described in U.S. Pat. No. 6,554,822, which is
incorporated herein by reference. External control can be initiated
by a caregiver. Alternatively, a subject can initiate delivery of
one or more label. As such, the system can have a built in
mechanism to limit the number of allowable doses by a vertebrate
subject and/or caregiver in a given time frame as described, for
example, in U.S. Pat. No. 6,796,956, which is incorporated herein
by reference.
Device Including Energy Sources for Eliciting One or More Signal
Responses
[0127] The device can include one or more energy sources configured
to provide energy to elicit one or more signal responses associated
with one or more labeled target components. In an aspect, the one
or more energy sources can be configured to elicit one or more
signal responses including, but not limited to, electromagnetic
signal responses, for example, optical signal responses (e.g.,
visible light signal responses, infrared signal responses,
ultraviolet signal responses, and/or fluorescent signal responses),
radiofrequency signal responses, and/or magnetic signal responses.
In another aspect, the one or more energy sources can be configured
to elicit one or more signal responses including a scattering
response and/or an absorptive response from the one or more target
components. In an aspect, the one or more signal responses can
include, but are not limited to, acoustic signal responses,
including, but not limited to, ultrasonic signal responses. One or
more signal responses can include, but are not limited to, thermal
signal responses, and/or color signal responses. Characteristics of
the energy appropriate for generating one or more signal responses
are known in the art and/or described herein.
[0128] Electromagnetic energy can include, but is not limited to,
radio waves, microwaves, terahertz radiation, infrared radiation,
visible light, X-rays, and gamma rays. In an aspect, one or more of
these frequencies can be explicitly excluded from the general
category of electromagnetic energy (e.g. electromagnetic energy
sources, but not including X-ray energy sources). Electromagnetic
energy, (or radiation) with a wavelength between approximately 400
nm and 700 nm is detected by the human eye and perceived as visible
light. Optical light can also include near infrared (longer than
700 nm) and ultraviolet (shorter than 400 nm).
[0129] Electromagnetic or optical energy is made up of photons and
can include single photon electromagnetic energy, two photon
electromagnetic energy, multiple wavelength electromagnetic energy,
and extended-spectrum electromagnetic energy. In an aspect, the
electromagnetic energy is generated by two photons having the same
wavelength. In an aspect, the electromagnetic energy is generated
by two photons having a different wavelength. Electromagnetic
energy generated by two photons is optionally focused at a depth
within a vessel lumen and/or a reservoir, optionally at one or more
depths. "Two-photon" energy generation can include excitation of a
fluorophore by two photons in a quantum event, resulting in the
emission of a fluorescence photon, optionally at a higher energy
than either of the two excitatory photons, optionally using a
femtosecond laser. In an aspect, two photon electromagnetic energy
is coupled through a virtual energy level and/or coupled through an
intermediate energy level. "Extended-spectrum" can include a range
of possible electromagnetic radiation wavelengths within the full
spectrum of possible wavelengths, optionally from extremely long to
extremely short.
[0130] Electromagnetic energy can be used to induce fluorescence,
which includes production of light (emission) following excitation
by electromagnetic energy. In an aspect, fluorescence can be
emitted from one or more labels bound to one or more target
components in response to an excitation energy. In an aspect,
inherent fluorescence, e.g., autofluorescence, can be emitted from
a target component in response to an excitation energy. In some
instances, fluorescence from a label and inherent autofluorescence
can be used in combination to generate signal responses associated
with one or more labeled target components.
[0131] Electromagnetic energy sources can be configured to emit
energy as a continuous beam or as a train of short pulses. In the
continuous wave mode of operation, the output is relatively
consistent with respect to time. In the pulsed mode of operation,
the output varies with respect to time, optionally having
alternating "on" and "off" periods. In illustrative examples, one
or more energy sources can be configured to emit continuous energy
to excite fluorophore-based labels to emit fluorescence. In an
aspect, the one or more electromagnetic energy sources include one
or more lasers having one or more of a continuous or pulsed mode of
action. One or more pulsed lasers can include, but are not limited
to, Q-switched lasers, mode locking lasers, and pulsed-pumping
lasers. Mode locked lasers emit extremely short pulses on the order
of tens of picoseconds down to less than 10 femtoseconds, the
pulses optionally separated by the time that a pulse takes to
complete one round trip in the resonator cavity. Due to the Fourier
limit, a pulse of such short temporal length can have a spectrum
which contains a wide range of wavelengths.
[0132] In an aspect, the energy, optionally electromagnetic energy,
can be defined spatially and/or directionally. In an aspect, the
electromagnetic energy can be spatially limited, optionally
spatially focused and/or spatially collimated. The electromagnetic
energy optionally contacts less than an entire possible area, or an
entire possible target, and/or is limited to a certain depth within
at least one lumen, and/or a reservoir.
[0133] In an aspect, the energy, optionally electromagnetic energy,
can be directionally limited, directionally varied, and/or
directionally variable. In an aspect, the energy can be provided
only in a single direction, for example 90 degrees from the
horizontal axis of a device, or toward a lumen wall, a bypass wall,
and/or a reservoir wall. In an aspect, the energy can be provided
over a range of directions for example, through movement of the
energy source, through movement of the entire device, and/or
through illumination from a variety of energy sources in the
device.
[0134] In an aspect, the one or more energy sources can be an
acoustic energy source configured to induce a signal response. The
one or more acoustic energy source can elicit one or more of a
signal response using ultrasound imaging. As such, a sound wave can
be sent through a piece of quartz or glass coated with a thin layer
of piezoelectric material that resonates at a specific frequency,
for example, 1 GHz, and through a lens to scan a cell or cells.
See, e.g., Ouellette. The Industrial Physicist Jun./Jul. 14-17,
2004, which is incorporated herein by reference. The sound waves
are reflected back up through the lens and piezoelectric material
which serve as detector and amplifier. The reflected sound waves
are recorded electronically and can be used directly to compare
cellular profiles. Alternatively, the recorded sound waves can be
converted into an image.
[0135] The resolution of acoustic imaging is dependent upon the
frequency of sound used for imaging. Standard ultrasound imaging
uses sound waves ranging in frequency from 3-10 MHz, but does not
provide cellular detail. Higher frequency ultrasound in the range
of 20-100 MHz can be used to detect changes in cellular structures
in tissues and cells, although individual cells are still not
easily resolved. For example, cells undergoing mitosis and cells
undergoing apoptosis in response to a chemotherapeutic agent, for
example, exhibit increased backscattered signal relative to normal
cells. See, e.g., Baddour et al. Ultrasonics Symposium IEEE
2:1639-1644, 2002, which is incorporated herein by reference.
Cellular resolution can be attained using sound waves ranging in
frequency from 100 MHz to 2 GHz, comparable to the range used for
acoustic microscopy. See, e.g., Schenk et al. J. Histochem.
Cytochem. 36:1341-1351, 1988, which is incorporated herein by
reference. The latter corresponds to wavelengths of 15 to 0.75
microns in water, the medium through which ultrasound and acoustic
imaging are done.
[0136] Photoacoustic imaging, in which ultrasound detection is
combined with optical stimulation, can also be used as one or more
energy sources to elicit one or more signal responses associated
with one or more labeled target components. See, e.g. Wygant et al.
IEEE Ultrasonics Symposium 1921-1924, 2005, which is incorporated
herein by reference. In this process, the optical absorption
properties of a material are imaged by detecting the ultrasound
emitted when a cell is illuminated with a laser. The emitted
ultrasound is due to the brief thermal expansions that occur when
the laser energy is absorbed by the cell. Those regions that are
more optically absorbent will generate a stronger acoustic signal.
Laser pulse widths, for example, of 10 ns and wavelengths between
600 nm and 1000 nm can be used for photoacoustic imaging of cells.
A single mechanically scanned piezoelectric transducer or a
capacitive micromachined ultrasonic transducer array, for example,
can be used to detect the laser generated ultrasound. See, e.g.
Wygant et al. IEEE Ultrasonics Symposium 1921-1924, 2005, which is
incorporated herein by reference.
[0137] Photoacoustic imaging can also be used to detect flowing
cells in vivo. A cell or cells can be irradiated with one or
several focused electromagnetic energy operating at different
wavelengths ranging, for example, from 415 to 2300 nm. An
ultrasound transducer can be used to record light-induced acoustic
waves. See, e.g. Zharov et al. SPIE Newsroom
10.1117/2.1200609.0391, 2006, which is incorporated herein by
reference.
[0138] Electromagnetic energy and/or acoustic energy configured to
induce a signal response in a target can be selected, optionally
manually, remotely, programmably, wirelessly, and/or using feedback
information. Frequencies that induce a signal response in one or
more targets are known in the art and/or discussed herein. In an
aspect, selection of excitation energy can be performed in advance,
or as a result of information received, optionally including
feedback information, optionally from one or more sensors or
provided by one or more external sources.
[0139] In an aspect, the at least one energy source can be an
electrical energy source configured to induce a signal response.
The one or more electrical energy sources can elicit one or more of
a signal response that is a change in electrical impedance. The
change in electrical impedance can be used to assess the relative
size of one or more target component. For example, a portion of the
lumen can include two or more electrodes between which an electric
current flows. A target component, e.g., target cell, passing
through the electric current displaces its own volume of conducting
fluid (i.e., plasma), momentarily increasing the impedance and
inducing a current fluctuation that can be converted into a voltage
pulse. The amplitude of the pulse is directly proportional to the
volume of the cell that produced it. As such, the device can be
configured to detect changes in electrical impedance to generate
volume information that can be compared with preset algorithms
defining, for example, the volume of normal blood cells,
pathological blood cells, and/or cells foreign to the circulation
such as pathogen or circulating tumor cells.
[0140] Target components of the blood or lymph can be
differentiated based on a volume using electrical impedance as
described herein and/or commonly practiced using a Coulter counter.
For example, platelets range in volume from 2 to 20 femtoliters
(fL) whereas red blood cells range in volume from 70-90 fL.
Neutrophils and eosinophils range in volume from 160 to 450 fL
while monocytes range in volume from 90 to 160 fL. By comparison,
bacteria may be as small as 1 fL whereas a circulating tumor cell
may be as large as 2000 fL. A MEMS resembling a miniaturized
Coulter counter can be incorporated into the device described
herein and can be constructed using thin platinum electrodes with a
sensing zone of, for example, 20-100 microns. See, e.g., Zheng et
al. Proceedings of 2006 International Conference on
Microtechnologies in Medicine and Biology, IEEE, Okinawa, Japan,
9-12 May, 2006; Gao et al. Proceedings of the 25.sup.th Annual
International Conference of the IEEE EMBS, Cancun, Mexico, Sep.
17-21, 2003, each of which is incorporated herein by reference.
[0141] Energy sources include, but are not limited to,
light-emitting diodes (LED), infrared LED, laser diodes (infrared
or near infrared), laser source (Argon-, HeNe-, GaAlAs-,
Nd:YAG-type lasers) directed heat source, electromagnetic energy
source,
[0142] The electromagnetic energy source can be one or more laser
diode placed in or proximal to the lumen of the blood vessel and
capable of producing energy sufficient to excite the
fluorescently-labeled metastatic tumor cells. The one or more laser
diode can produce energy in the 750-850 nm range that is
configured, if needed, to penetrate the blood vessel wall. Near
infrared light at 800 nm can at least partially penetrate through 7
millimeters of vertebrate tissue. See, e.g., Fischer, et al.,
Conference on Lasers and Electro-Optics Europe; CLEO/Europe, 2005;
Publication date Jun. 17, 2005, pp. 641, which is incorporated
herein by reference. Optionally, wavelengths in a wider range such
as 720-904 nm can be used. Laser diodes (3-9 millimeters in
diameter) emitting in the far red and near infrared wavelengths are
commercially available (from, e.g., JDS Uniphase Corporation,
Milpitas, Calif.). Alternatively, the energy source can be one or
more far red or near infrared LED chip available in sub-millimeter
dimensions (from, e.g., Marubeni America Corporation, Santa Clara,
Calif.). The wavelength used by the device depends on the
absorbance/emission properties of the label and the tissue
penetration depth that is required. Other examples of light or
energy sources include, but are not limited to, Argon-, HeNe-,
GaAlAs-, Nd:YAG-type lasers.
Device Including Sensors Configured to Detect Signal Responses
[0143] The device can include one or more sensors configured to
detect one or more signal responses associated with the one or more
labeled target components. One or more imaging sensors can include,
but are not limited to, one or more electromagnetic energy sensors
and/or one or more acoustic sensors, e.g. ultrasonic sensors. One
or more electromagnetic energy sensors can include, but are not
limited to, one or more photodetectors, one or more radiofrequency
antennae, one or more magnetic energy sensors, one or more thermal
sensors, and/or one or more electrical energy sensors. One or more
electromagnetic energy sensors can include one or more optical
sensors including, but not limited to, sensors configured to detect
near IR, UV, fluorescence, and/or visual light.
[0144] One or more imaging sensors can include, but are not limited
to, photodiodes, photoresistors, charged-coupled devices (CCD)
and/or complementary metal oxide semiconductor (CMOS) cameras. One
or more imaging sensors can include, but are not limited to, one or
more piezo transducers, one or more MEMS devices, one or more
cavity resonators, one or more magneto resistive sensors, one or
more magnetic field sensors, and/or one or more thermal sensors. An
example of an implantable CMOS imaging sensor for use in imaging a
vertebrate subject in vivo is described in Tamura, et al. J.
Neurosci. Methods 173:114-120, 2008, which is incorporated herein
by reference.
[0145] In an aspect, the one or more sensors can be configured to
detect one or more signal responses associated with one or more
labeled target components that are one or more labeled cells in the
blood or lymph of a vertebrate subject. A labeled target component
that is a labeled cell can include, for example, a bacterium, a
protozoan, a platelet, a red blood cell, a lymphocyte, a monocyte,
a neutrophil, an eosinophil, a circulating tumor cell, or a
combination there of. A signal response associated with one or more
labeled cell can be detected using any of a number of imaging or
optical methods including, but not limited to, light scattering,
electrical impedance, infrared spectroscopy, acoustic imaging,
thermal imaging, photothermal imaging, dark field, visible light
absorption and refraction, and autofluorescence. See, e.g., U.S.
Patent Application 2009/0093728; Doornbos et al. Cytometry
14:589-594, 1993; Gao et al. Proceedings of the 25.sup.th Annual
International Conference of the IEEE EMBS, Cancun, Mexico, Sep.
17-21, 2003; Oberreuter et al. Int. J. Syst. Evol. Microbiol.
52:91-100, 2002; Baddour et al. Ultrasonics Symposium IEEE
2:1639-1644, 2002; Zharov et al. J. Cell. Biochem. 97:916-932,
2006; Zharov et al. J. Biomed. Opt. 11:054034-1-4, 2006; Koenig et
al. J. Fluoresc. 4:17-40, 1994; each of which is incorporated
herein by reference. For example, red blood cells infected with the
parasite Plasmodium falciparum can be differentiated from other
cells in the blood using differential light scatter at 10 degrees
(complexity) and polarized light scatter at 90 degrees (lobularity)
based on the pigmentation of the parasite. See, e.g., Mendelow et
al. Br. J. Haematology 104:499-503, 1999, which is incorporated
herein by reference.
[0146] In some instances, one or more target cells in the blood or
lymph of a vertebrate subject can be recognized based on a spectral
analysis of the target cells. Alternatively, the one or more target
cells can be recognized based on pattern and image recognition or
signal recognition analysis. Various methods have been described
for image and shape analysis of cells and subcellular components of
cells. See, e.g., U.S. Pat. Nos. 5,107,422; 5,790,691; 6,956,961
B2; 7,151,847 B2; U.S. Patent Applications 2005/0251347 A1;
2006/0039593 A1; Fei-Fei et al. IEEE Transactions on Pattern
Analysis and Machine Intelligence 28:594-611, 2006; Martin et al.
IEEE Transactions on Pattern Analysis and Machine Intelligence
26:530-549, 2004; Olson et al. Proc. Natl. Acad. Sci. USA
77:1516-1520, 1980; Schneider, et al Biorheology 32:237-238, 1995;
each of which is incorporated herein by reference. For example, a
Texture Analyzing System can be used to distinguish various cells
in the blood and/or lymph of a vertebrate subject based on the
granularity of the cell or cells. See, e.g., Bins et al. Cytometry
1:321-324, 1981, which is incorporated herein by reference. The
imaged components of the cells are measured with a gray scale with
33 intervals ranging from black (level 0) to white (level 99) and a
histogram is generated. Mature white blood cells (neutrophils,
eosinophils, basophils and lymphocytes) have a dense nuclear
structure and therefore low counts. In contrast, monocytes have a
looser, less dense nuclear structure and high counts. The cytoplasm
of eosinophils and neutrophils is very granular and is reflected in
the combination of high positive and low negative counts. Smaller
values are seen in the cytoplasm of lymphocytes, monocytes and
basophils. Similarly, granulometries can be used to identify red
blood cells infected with the malarial parasite. See, e.g.,
Dempster & DiRuberto Circuits and Systems, 2001, ISCAS 2001,
The 2001 IEEE International Symposium 5: 291-294, May 6-9, 2001,
which is incorporated herein by reference.
[0147] In an aspect, one or more sensors can be configured to
capture one or more signal responses including, but not limited to,
electromagnetic signal responses including, but not limited to,
optical signal responses (e.g. visible light signal responses,
infrared signal responses, ultraviolet signal responses, and/or
fluorescent signal responses, among others), radiofrequency signal
responses, and/or magnetic signal responses. One or more signal
responses can include, but are not limited to, acoustic signal
responses, including, but not limited to, ultrasonic signal
responses. One or more signal responses can include, but are not
limited to, thermal signal responses, and/or color signal
responses. In an aspect, one or more sensors are configured to
capture one or more signal responses at one or more wavelengths. In
an aspect, one or more signal responses described herein and/or
known in the art may be specifically excluded from an embodiment,
e.g. signal responses not including fluorescent signal
responses.
[0148] In an aspect, one or more signal responses can include, but
are not limited to, actual images of cells or particles within the
vessel lumen. In an aspect, one or more signal responses include
image information such as cell or particle shape, cell or particle
outline, and/or cell or particle periphery, among others. In an
aspect, one or more signal responses include image information such
as intracellular shapes, intracellular outlines, and/or
intracellular peripheries, among others. In an aspect, one or more
sensors can be configured to capture light scattering. In an
aspect, one or more sensors can be configured to capture fluid
and/or cell or particle velocity.
[0149] One or more electromagnetic energy sensors can be configured
to measure the absorption, emission, fluorescence, or
phosphorescence of one or more labeled target components. Such
electromagnetic properties can be inherent properties of all or a
portion of one or more target components (e.g. auto-fluorescence),
or can be associated with one or more labels added or introduced to
the body, surface, lumen, interior, and/or fluid and configured to
bind to one or more targets. One or more target components can
include, but are not limited to, one or more cells.
[0150] In an aspect, one or more sensors can be configured to
detect a fluorescent response at a single wavelength of
electromagnetic energy, at two wavelengths of electromagnetic
energy, at multiple wavelengths of electromagnetic energy, or over
extended-spectrum electromagnetic energy. In an aspect, one or more
sensors can be configured to detect excitation energy.
[0151] In an aspect, one or more sensors can be configured to
detect a cumulative (optionally fluorescent) response over a time
interval. In an aspect, one or more sensors can be configured to
detect a (optionally fluorescent) response at a specific time
interval and/or at a specific time. In an aspect, one or more
sensors are configured to detect a time-dependent (optionally
fluorescent) response. In illustrative examples, the cumulative
response is determined over milliseconds, seconds, and/or minutes
following excitation. In an aspect, the response is detected over
millisecond, second, and/or minute time intervals following
excitation. In an, aspect, the response is detected approximately
femtoseconds, picoseconds, nanoseconds, milliseconds, seconds,
and/or minutes after excitation.
[0152] In an aspect, one or more sensors can be configured to be
calibrated optionally at least partially based an expected baseline
signal response, e.g. normal signal response, for the fluid,
tissue, cells, and/or lumen. "Normal signal response" can include
the detected intrinsic signal response of one or more fluid,
tissue, cells, and/or lumen as determined by researchers and/or
medical or veterinary professionals for subjects of a certain age
or ethnicity who do not have pathological conditions (e.g. control
subjects). Normal signal response can include the intrinsic
detected signal response of fluid, tissue, cells, and/or lumen of a
vertebrate subject prior to a pathological condition and/or of a
comparable location not affected by the pathological condition.
[0153] In an aspect, the device can include one or more sensors
that can be configured to detect a condition of interest including,
but not limited to, a temperature, a pressure, a fluid flow, an
optical absorption, optical emission, fluorescence, or
phosphorescence, an index of refraction at least one wavelength, an
electrical resistivity, a density or sound speed, a pH, an
osmolality, the presence of an embolism, the presence or absence of
an object, for example, a blood clot, a thrombus, an embolus, a
plaque, a lipid, a gas bubble, an aggregate, a cell, a specific
type of cell, a cellular component or fragment, a collection of
cell, a gamete, a pathogen, or a parasite, and/or the presence or
absence of a substance, for example, a biological marker, an
antibody, an antigen, a peptide, a polypeptide, a protein, a
complex, a nucleic acid, a cell and/or a cell of a particular type,
a cellular component, an organelle, a pathogen, a lipid, a
lipoprotein, an alcohol, an acid, an ion, an immunomodulator, a
sterol, a carbohydrate, a polysaccharide, a glycoprotein, a metal,
an electrolyte, a metabolite, an organic compound, an
organophosphate, a drug, a therapeutic, a gas, and/or a
pollutant.
[0154] In an aspect, the one or more sensors can be radioactivity
sensors configured to detect radioactivity associated with a
radioactive label bound to one or more target components. Examples
of methods for detecting radioactivity include, but are not limited
to, gas-filled tube detectors, e.g., Geiger counters; scintillation
crystal detectors; and solid-state semiconductor detectors. In an
aspect, the one or more radioactivity sensors are one or more of a
scintillation crystal detector made from a material that fluoresces
or emits light when hit by radiation particles. The fluorescence or
light can be measured using one or more of a light capture device,
e.g., a photomultiplier or photodiode. In an aspect, the one or
more radioactivity sensors are one or more of a microdosimeter. A
microdosimeter for wireless measurement of radioactivity in vivo
has been described and is configured to detect ionizing radiation
using a parallel plate capacitor to form a passive LC resonator.
See, e.g., Son & Ziaie. IEEE Trans. Biomed Eng. 55:1772-1775,
2008, which is incorporated herein by reference.
[0155] In an aspect, the one or more sensors can be one or more
magnetic sensors capable of detecting a target component labeled
with a magnetic particle. Examples of MEMS magnetic sensors are
described in Lee, et al. Magnetics Conference, 2006. INTERMAG 2006.
IEEE International; Erye et al., "MEMS Magnetic Sensor in Standard
CMOS", in Science Closure and Enabling Technologies for
Constellation Class Mission, ed. V. Anelopoulos and P. V. Panetta,
pp. 99-102, UC Berkeley, Calif., 1998, each of each of which is
incorporated herein by reference.
Device Including Additional Sensors
[0156] The device can include one or more additional sensors
configured to sense target components directly and/or to sense
disease-associated physiological parameters. For example, the
device can include one or more physiological sensors configured to
detect one or more of a body temperature, pH, pressure, edema,
oxygen level, or toxin level. One or more physiological sensors can
include, but are not limited to, one or more chemical and/or
biological molecule sensors, e.g. blood chemistry, chemical
concentration, biosensors; one or more pH sensors; one or more time
sensors, e.g. timers, clocks; and/or one or more temperature
sensors. One or more physiological detectors can include, but are
not limited to, blood pressure detectors, pulse detectors,
peristaltic action sensors, pressure sensors, flow sensors,
viscosity sensors, and/or shear sensors.
[0157] The device including one or more sensors can be configured
to measure various parameters including, but not limited to, the
electrical resistivity of the fluid, the density or sound speed of
the fluid, the pH, the osmolality, or the index of refraction of
the fluid at least one wavelength. The selection of a suitable
sensor for a particular application or use site is considered to be
within the capability of a person having skill in the art. One or
more of these and/or other sensing capabilities can be present in a
single sensor or an array of sensors; sensing capabilities are not
limited to a particular number or type of sensors.
[0158] One or more biosensors can be configured to detect materials
including, but not limited to, a biological marker, an antibody, an
antigen, a peptide, a polypeptide, a protein, a complex, a nucleic
acid, a cell and, in some cases, a cell of a particular type, e.g.,
by methods used in flow cytometry. One or more biosensors can be
configured to detect materials including, but not limited to, a
cellular component, an organelle, a gamete, a pathogen, a lipid, a
lipoprotein, an alcohol, an acid, an ion, an immunomodulator, a
sterol, a carbohydrate, a polysaccharide, a glycoprotein, a metal,
an electrolyte, a metabolite, an organic compound, an
organophosphate, a drug, a therapeutic, a gas, a pollutant, or a
tag. A biosensor can include an antibody or other binding molecule
such as a receptor or ligand.
[0159] One or more sensors can include a gas sensor such as an
acoustic wave, chemiresistant, or piezoelectric sensors, or an
electronic nose. One or more sensors are optionally small in size,
for example a sensor or array that is a chemical sensor (Snow,
Science 307: 1942-1945, 2005), a gas sensor (Hagleitner, et al.,
Nature 414: 293-296, 2001.), an electronic nose, and/or a nuclear
magnetic resonance imager (Yusa (2005), Nature 434:1001-1005).
Additional examples of sensors are provided in The Biomedical
Engineering Handbook, Second Edition, Volume I, J. D. Bronzino,
Ed., Copyright 2000, CRC Press LLC, pp. V-1-51-9, and U.S. Pat. No.
6,802,811), each of which is incorporated herein by reference.
[0160] The device can include one or more sensors for qualitatively
and/or quantitatively measuring one or more target components in
the blood of a vertebrate subject. The one or more sensors can
include, but are not limited to, one or more of a biosensor, a
chemical sensor, a physical sensor, an optical sensor, or a
combination thereof. The one or more sensors can include one or
more target recognition elements that recognize one or more target
components. The interaction of one or more target components with
one or more sensors results in one or more detectable signals sent
to the controller.
[0161] In another aspect, the device including one or more sensors
can include one or more target recognition elements or labels that
recognize one or more target components. The target recognition
elements or labels are configured to specifically bind one or more
target components and can include, but are not limited to,
antibodies, antibody fragments, peptides, oligonucleotides, DNA,
RNA, aptamers, protein nucleic acids, proteins, viruses, enzymes,
receptors, bacteria, cells, cell fragments, inorganic molecules,
organic molecules, or combinations thereof. The one or more target
recognition elements or labels can be associated with one or more
substrate integrated into the one or more sensors. Binding of a
target component to a specific target recognition element or label
activates the sensor.
[0162] In an aspect, the one or more sensors can use Forster or
fluorescence resonance energy transfer (FRET) to sense one or more
target components in the blood or lymph of a vertebrate subject.
FRET is a distance-dependent interaction between the electronic
excited states of two dye molecules in which excitation is
transferred from a donor molecule to an acceptor molecule without
emission of a photon. For use in a sensor, the one or more target
recognition elements or labels associated with the one or more
sensors can include at least one donor molecule and at least one
acceptor molecule. Binding of a target component to the target
recognition element or label can result in a conformation change in
the target recognition element or label, leading to changes in the
distance between the donor and acceptor molecules and changes in
measurable fluorescence.
[0163] A variety of donor and acceptor fluorophore pairs can be
considered for FRET associated with the target recognition element
or label including, but not limited to, fluorescein and
tetramethylrhodamine; IAEDANS and fluorescein; fluorescein and
fluorescein; and BODIPY FL and BODIPY FL, and various Alexa Fluor
pairings as described herein. The cyanine dyes Cy3, Cy5, Cy5.5 and
Cy7, which emit in the red and far red wavelength range (>550
nm) as well as semiconductor quantum dots can also be used for
FRET-based detection systems. Quenching dyes can also be used to
quench the fluorescence of visible light-excited fluorophores,
examples of which include DABCYL, the non-fluorescing
diarylrhodamine derivative dyes QSY 7, QSY 9 and QSY 21 (Molecular
Probes, Carlsbad, Calif., USA), the non-fluorescing Black Hole
Quenchers BHQ0, BHQ1, BHQ2, and BHQ3 (Biosearch Technologies, Inc.,
Novato, Calif., USA) and Eclipse (Applera Corp., Norwalk, Conn.,
USA). A variety of donor fluorophore and quencher pairs can be
considered for FRET associated with the target recognition element
including, but not limited to, fluorescein with DABCYL; EDANS with
DABCYL; or fluorescein with QSY 7 and QSY 9. In general, QSY 7 and
QSY 9 dyes efficiently quench the fluorescence emission of donor
dyes including blue-fluorescent coumarins, green- or
orange-fluorescent dyes, and conjugates of the Texas Red and Alexa
Fluor 594 dyes. QSY 21 dye efficiently quenches all red-fluorescent
dyes. A number of the Alexa Fluor (AF) fluorophores (Molecular
Probes-Invitrogen, Carlsbad, Calif., USA) can be paired with
quenching molecules as follows: AF 350 with QSY 35 or DABCYL; AF
488 with QSY 35, DABCYL, QSY7 or QSY9; AF 546 with QSY 35, DABCYL,
QSY7 or QSY9; AF 555 with QSY7 or QSY9; AF 568 with QSY7, QSY9 or
QSY21; AF 594 with QSY21; and AF 647 with QSY 21.
[0164] The device including one or more sensors can sense the
target components in the blood or lymph of a vertebrate subject
using the technique of surface plasmon resonance (for planar
surfaces) or localized surface plasmon resonance (for
nanoparticles). Surface plasmon resonance involves detecting
changes in the refractive index on a sensor surface in response to
changes in molecules bound on the sensor surface. In an aspect, the
surface of the sensor is a glass support or other solid support
coated with a thin film of metal, for example, gold. In an aspect,
the sensor surface includes a matrix to which is immobilized one or
more target recognition elements that recognize one or more target
components. The target recognition elements or labels can be
antibodies or fragments thereof, oligonucleotide or peptide based
aptamers, receptors or ligands, artificial binding substrates
formed by molecular imprinting, or any other examples of molecules
and/or substrates that bind cells. As blood or lymph from the
vertebrate subject passes by the sensor surface, a target component
can interact with a target recognition element or label on the
sensor surface. The sensor is illuminated by monochromatic light.
Resonance occurs at a specific angle of incident light. The
resonance angle depends on the refractive index in the vicinity of
the surface, which is dependent upon the concentration of target
components on the surface. An example of instrumentation that uses
surface plasmon resonance is the BIACORE system (Biacore, Inc.--GE
Healthcare, Piscataway, N.J.) which includes a sensor microchip, a
laser light source emitting polarized light, an automated fluid
handling system, and a diode array position sensitive detector.
See, e.g., Raghavan & Bjorkman Structure 3:331-333, 1995, which
is incorporated herein by reference.
[0165] The device including one or more sensors for sensing target
components can include one or more label-free optical biosensors
that incorporate other optical methodologies, e.g.,
interferometers, waveguides, fiber gratings, ring resonators, and
photonic crystals. See, e.g., Fan, et al., Anal. Chim. Acta
620:8-26, 2008, which is incorporated herein by reference.
[0166] The device including one or more sensors for sensing target
components can include one or more microcantilevers. A
microcantilever can act as a biological sensor by detecting changes
in cantilever bending or vibrational frequency in response to
binding of one or more target components to the surface of the
sensor. In an aspect the sensor can be bound to a microcantilever
or a microbead as in an immunoaffinity binding array. In another
aspect, a biochip can be formed that uses microcantilever
bi-material formed from gold and silicon, as sensing elements. See,
e.g. Vashist J. Nanotech Online 3:DO: 10.2240/azojono0115, 2007,
which is incorporated herein by reference. The gold component of
the microcantilever can be coated with one or more target
recognition elements or labels which upon binding one or more
target components cause the microcantilever to deflect. Aptamers or
antibodies specific for one or more target components can be used
to coat microcantilevers. See, e.g., U.S. Pat. No. 7,097,662, which
is incorporated herein by reference. The one or more sensor can
incorporate one or more methods for microcantilever deflection
detection including, but not limited to, piezoresistive deflection,
optical deflection, capacitive deflection, interferometry
deflection, optical diffraction grating deflection, and charge
coupled device. In an aspect, the one or more microcantilever can
be a nanocantilever with nanoscale components. The one or more
microcantilevers and/or nanocantilevers can be arranged into
arrays. Both microcantilevers and nanocantilevers can find utility
in microelectomechnical systems (MEMS) and/or nanoelectomechnical
systems (NEMS).
[0167] The device including one or more sensors for sensing target
components can include a field effect transistor (FET) based
biosensor. In an aspect, a change in electrical signal is used to
detect interaction of one or more target components with one or
more target recognition elements of the sensor. See, e.g., U.S.
Pat. No. 7,303,875, which is incorporated herein by reference.
[0168] The device including one or more sensors for sensing one or
more target components can incorporate electrochemical impedance
spectroscopy. Electrochemical impedance spectroscopy can be used to
measure impedance across a natural and/or artificial lipid bilayer.
The sensor can incorporate an artificial bilayer that is tethered
to the surface of a solid electrode. One or more receptor can be
embedded into the lipid bilayer. The one or more receptors can be
ion channels that open and close in response to binding of a
specific analyte. The open and closed states can be quantitatively
measured as changes in impedance across the lipid bilayer. See,
e.g., Yang, et al., IEEE SENSORS 2006, EXCO, Daegu, Korea/Oct.
22-25, 2006, which is incorporated herein by reference.
[0169] The device including one or more sensors can include cells
with binding elements which when bound to target components induce
a measurable or detectable change in the cells. The cells can emit
a fluorescent signal in response to interacting with one or target
components. For example, a bioluminescent bioreporter integrated
circuit can be used in which binding of a ligand to a cell induces
expression of reporter polypeptide linked to a luminescent
response. See, e.g., U.S. Pat. No. 6,673,596; Durick &
Negulescu Biosens. Bioelectron. 16:587-592, 2001; each of which is
incorporated herein by reference. Alternatively, the one or more
cells can emit an electrical signal in response to interacting with
one or more target components. In an aspect, an implantable
biosensor can be used which is composed of genetically-modified
cells that respond to target binding by emitting a measurable
electrical signal. See U.S. Patent Application 2006/0234369 A1;
which is incorporated herein by reference.
[0170] The target recognition elements or labels recognized by the
one or more sensors can detect one or more target components
described herein and broadly including, but not limited to,
non-cellular target components such as sugars, lipids, vitamins,
minerals, non-protein hormones, proteins, serum proteins, acute
phase proteins associated with disease, coagulation or complement
related proteins, markers of cellular activation, soluble
inflammatory mediators, legal drugs, illicit drugs, and
environmental toxins; and cellular target components such as blood
cells (e.g., red blood cells, platelets, lymphocytes, monocytes,
neutrophils, eosinophils, basophils), viruses (e.g., human
immunodeficiency virus (HIV) and the hepatitis B, hepatitis C, and
hepatitis D viruses), bacteria (e.g., Staphylococcus,
Streptococcus, Pseudomonas, Haemophilus, Listeria, Esherichia
coli), fungi, (e.g., Candida albicans, Candida glabrata,
Aspergillus, T. glabrata, Candida tropicalis, C. krusei, and C.
parapsilosis) parasites (e.g., Trypanosoma cruzi, Trypanosoma
brucei, Leishmania, Plasmodium, Babesia microti, Toxoplasma gondii)
and cancer cells (e.g., metastatic tumor cells, hematopoietic
cancer cells).
[0171] The one or more target recognition elements or labels can be
configured to recognize one or more biomolecules on the surface of
the one or more target cells. In an aspect, the one or more target
recognition elements, e.g., antibody, aptamer, ligand, or ligand
mimetic, can be configured to recognize one or more receptor types
on the surface of target cells. Examples of receptors include, but
are not limited to, acetylcholine receptors, adenosine receptors,
adrenoceptors, GABA receptors, angiotensin receptors, cannabinoid
receptors, cholecystokinin receptors, dopamine receptors, glucagon
receptors, glucocorticoid receptors, glutamate receptors, histamine
receptors, mineralocorticoid receptors, olfactory receptors, opioid
receptors, purinergic receptors, secretin receptors, serotonin
receptors, somatostatin receptors, steroid hormone receptors,
calcium-sensing receptor, hormone receptors, erythropoietin
receptor, and natriuretic peptide receptors. Other examples include
type I cytokine receptors (e.g., type I interleukin receptors,
erythropoietin receptor, GM-CSF receptor, G-CSF receptor, growth
hormone receptor, oncostatin M receptor, leukemia inhibitory factor
receptor); type II cytokine receptors (e.g., type II interleukin
receptors, interferon-.alpha./.beta. receptors, interferon-.gamma.
receptor); members of the immunoglobulin superfamily (e.g.,
interleukin-1 receptor, CSF1, c-kit receptor, interleukin-18
receptor); tumor necrosis factor (TNF) receptor family (e.g., TNF
receptor 1 (TNF-R1), TNF receptor 2 (TNF-R2), CD27, CD40, and
lymphotoxin .beta. receptor); chemokine receptors including
serpentine CCR and CXCR receptors (e.g., CCR1 and CXCR4, and
interleukin-8 receptor); TGF .beta. receptors. See Ozaki and
Leonard, J. Biol. Chem. 277:29355-29358, 2002, which is
incorporated herein by reference.
[0172] In an aspect, the one or more target recognition elements or
labels, e.g., antibody, aptamer, ligand, or ligand mimetic, can be
configured to recognize other biomolecules on the surface of target
cells including, but not limited to, various CD (cluster of
designation/cluster of differentiation) markers, intergrins, ion
channels, ATPases, cell adhesion molecules, integral membrane
glycoproteins, immunoglobulins, transporters. The one or more
target recognition elements can be configured to recognize
components of cell surface biomolecules including amino acid
sequence and oligosaccharide modifications.
[0173] In an aspect, the target recognition element or label, e.g.,
antibody, aptamer, ligand, or ligand mimetic, can be configured to
recognize a biomolecule associated with a tumor cell. Examples of
tumor associated components can include, but are not limited to,
BLyS receptor, carcinoembryonic antigen (CA-125), CD25, CD34, CD33
and CD123 (acute myeloid leukemia), CD20 (chronic lymphocytic
leukemia), CD19 and CD22 (acute lymphoblastic leukemia), CD30,
CD40, CD70, CD133, 57 kD cytokeratin, epithelial specific antigen,
extracellular matrix glycoprotein tenascin, Fas/CD95,
gastrin-releasing peptide-like receptors, hepatocyte specific
antigen, human gastric mucin, human milk fat globule, lymphatic
endothelial cell marker, matrix metalloproteinase 9, melan A,
melanoma marker, mesothelin, mucin glycoproteins (e.g., MUC1, MUC2,
MUC4, MUC5AC, MUC6), prostate specific antigen, prostatic acid
phosphatase, PTEN, renal cell carcinoma marker, RGD-peptide binding
integrins, sialyl Lewis A, six-transmembrane epithelial antigen of
the prostate (STEAP), TNF receptor, TRAIL receptor, tyrosinase,
villin. Other tumor associated antigens include, but are not
limited to, alpha fetoprotein, apolipoprotein D, clusterin,
chromogranin A, myeloperoxidase, MyoD1 myoglobin placental alkaline
phosphatase c-fos, homeobox genes.
[0174] In an aspect, the target recognition element or label, e.g.,
antibody, aptamer, ligand, or ligand mimetic, can be configured to
recognize a biomolecule associated with the surface of a pathogen,
e.g., bacteria, a virus, a fungus, or a parasite. The biomolecule
can be one or more components of the bacterial outer membrane, cell
wall, and/or cytoplasmic membrane. Examples of components
associated with the bacterial outer membrane of Gram-negative
bacteria include, but are not limited to, lipopolysaccaride and OMP
(outer membrane protein) porins, the latter of which are
exemplified by OmpC, OmpF and PhoP of E. coli. Examples of
components associated with the bacterial cell wall of both
Gram-positive and Gram-negative bacterial include, but are not
limited to, peptidoglycans polymers composed of an alternating
sequence of N-acetylglucoamine and N-acetyl-muraminic acid and
crosslinked by amino acids and amino acid derivatives. Examples of
components associated with the bacterial cytoplasmic membrane
include, but are not limited to, the MPA1-C (also called
polysaccharide copolymerase, PCP2a) family of proteins, the MPA2
family of proteins, and the ABC bacteriocin exporter accessory
protein (BEA) family of proteins. Other examples of components
associated with bacteria include, but are not limited to,
transporters, e.g., sugar porter (major facilitator superfamily),
amino-acid/polyamine/organocation (APC) superfamily, cation
diffusion facilitator, resistance-nodulation-division type
transporter, SecDF, calcium:cation antiporter, inorganic phosphate
transporter, monovalent cation:proton antiporter-1, monovalent
cation:proton antiporter-2, potassium transporter,
nucleobase:cation symporter-2, formate-nitrite transporter,
divalent anion:sodium symporter, ammonium transporter, and
multi-antimicrobial extrusion; channels, e.g., major intrinsic
protein, chloride channel, and metal ion transporter; and primary
active transporters, e.g., P-type ATPase, arsenite-antimonite
efflux, Type II secretory pathway (SecY), and sodium-transporting
carboxylic acid decarboxylase. A number of other potential
components associated with bacteria have been described in Chung,
et al., J. Bacteriology 183:1012-1021, 2001, which is incorporated
herein by reference.
[0175] In an aspect, the target recognition element or label can be
configured to recognize a biomolecule associated with a blood cell
infected with a pathogen. For example, red blood cells infected
with P. falciparum can be distinguished from normal red blood cells
by changes in surface protein expression including expression on
the red blood cell surface of the parasite derived protein P.
falciparum erythrocyte membrane protein (PfEMP1). See, e.g.,
Horata, et al., Malaria J. 8:184, 2009, which is incorporated
herein by reference.
Device Including One or More Reactive Components
[0176] A device is disclosed herein that includes at least one
second reservoir configured to include one or more reactive
components responsive to the sensor, wherein the one or more second
reservoirs are located in communication with, or located within,
the at least one lumen and include the one or more reactive
components configured to modulate a physiological effect of the one
or more target components in the one or more of blood or lymph of a
vertebrate subject. The one or more reactive components, e.g.,
reactive chemical, components, reactive biologic components or
reactive physical components, include, but are not limited to, one
or more of a denaturing agent, a degradative agent, a
cell-disrupting agent, a modulator, an apoptotic agent, a cytotoxic
agent, cytostatic agent, a chemotherapeutic agent, an antibody
toxin, or a combination thereof. The one or more reactive chemical
components or reactive biologic components configured to modulate a
physiological effect of the one or more target components can be
any of a number of chemical types or biologic types including, but
not limited to, a protein, a peptide, a small molecule, a chemical,
a toxin, an aptamer, or an inhibitory RNA, DNA, or other nucleic
acid. The one or more reactive components can be incorporated into
or released within one or more second reservoirs associated with
the device. Alternatively, the one or more reactive components can
be diffusible components released from a reservoir of the device
into the blood of the vertebrate subject. The one or more reactive
components can further include one or more reactive physical
components including, but not limited to, one or more of polymers,
imprinted polymers, or charged polymers. In an aspect, the one or
more reactive physical components can include a charged plastic
polymer surface configured to bind cells, tumor cells, emboli,
misfolded proteins, aggregated proteins, autoimmune antibodies,
infectious agents, or infected cells.
[0177] In an aspect, the reactive biologic component can be a
recombinant protein or peptide. The recombinant protein or peptide
can be generated exogenously and incorporated into one or more
second reservoirs of the device. In an aspect, the recombinant
protein or peptide can be generated by one or more cells
incorporated into the device. The one or more cells can be
genetically modified to synthesize and secrete the one or more
reactive biologic components. Cells that can be used for this
purpose include, but are not limited to, mammalian cells,
enucleated cells (e.g., erythrocytes), plants cells, insect cells,
bacteria, or yeast. DNA sequences corresponding to one or more
reactive biologic components are cloned into an appropriate cell
type using standard procedures with appropriate expression vectors
and transfection protocols. The genetically modified cells are
encapsulated in one or more compartments of the device and secrete
the one or more reactive biologic components into the blood or
lymph of a vertebrate subject. The genetically modified cells are
kept separate from the circulation of the vertebrate subject using
a size-limiting biocompatible mesh or membrane filter, for example,
that allows passage of the cytotoxic, cytostatic, apoptotic, and/or
chemotherapeutic agent, but not the larger cells.
[0178] In an aspect, the one or more reactive chemical components
or reactive biologic components can be released from synthetic
vesicles or particles. Examples include any of a number of drug
delivery vehicles including, but not limited to, phospholipid
vesicles, liposomes, nanoparticles, polymers, or hydrogels. The
release of the one or more reactive chemical component or reactive
biologic component can be triggered by binding of a specific target
to the synthetic vesicle or particle. For example, one or more DNA
aptamers can be incorporated into hydrogel and designed to bind one
or more specific targets and release the contents of the hydrogel
as described herein.
[0179] The device can include one or more reservoirs that store one
or more reactive components. The one or more reservoirs of the
device can be configured to controllably release one or more
reactive components. Each reservoir can contain one or more
reactive components. Release of the reactive component from a
reservoir is controlled by the controller component of the device.
In an aspect, the reactive components can be housed in multiple
reservoirs associated with the device. For example, the device can
include one or more microchips each with multiple reservoirs sealed
with removable caps to enable controlled release of one or more a
denaturing agent, a degradative agent, a cell-disrupting agent, a
modulator, an apoptotic agent, a cytotoxic agent, cytostatic agent,
a chemotherapeutic agent, an antibody toxin, reactive physical
components, or a combination thereof. See, e.g., U.S. Pat. No.
7,413,846; Maloney & Santini, Proceedings 26.sup.th Annual
International Conference IEEE EMBS, San Francisco, Calif., USA,
Sep. 1-5, 2004, which is incorporated herein by reference.
[0180] Binding agents remove one or more target components from the
blood. The device can include one or more reactive components that
are binding agents configured to remove one or more labeled target
components from the blood or lymph of a vertebrate subject. The one
or more binding agents can be used alone to selectively or
non-selectively bind and sequester one or more target components or
target cells from the blood or lymph of the vertebrate subject.
Alternatively, the one or more binding agents can be used to
capture one or more labeled target components in combination with a
treatment including one or more additional reactive components,
e.g., a second binding agent, a cell-disrupting agent, a cytotoxic
agent, a cytostatic agent, an apoptotic agent, a chemotherapeutic
agent, an antibody-toxin agent, or a combination thereof. Following
binding of the one or more labeled target components to the one or
more binding agents in at least one lumen, one or more additional
reactive components can be provided to modulate a physiological
effect of the one or more target components.
[0181] The one or more binding agents can be configured to
non-selectively bind one or more labeled target components. For
example, the binding agents can constitute all or part of one or
more components of extracellular matrix, e.g., fibronectin,
vitronectin, collagen, and laminin to which cells expressing
integrins and other cell surface components will non-selectively
bind. Alternatively, the binding agents can constitute all or part
of one or more components of basal lamina, e.g., collagen, heparan
sulfate proteoglycan, laminin, integrins, or dystroglycans, to
which cells expressing cell adhesion molecules and other cell
surface components will non-selectively bind. In an aspect, the
binding agent can be one or more of a commercially available cell
adhesion product (e.g., BD MATRIGEL.TM. from BD Biosciences, San
Jose, Calif.). In an aspect, the one or more binding agent can be a
surface substrate that non-selectively binds target cells, which
include, but are not limited to, glass or plastic.
[0182] The one or more binding agents can include absorbent
material that non-selectively binds one or more labeled target
component. The absorbent material can include, but is not limited
to, silica, activated charcoal, nonionic or uncharged resins or
polymers, ionic or charged resins or polymers, anion exchange
resins or polymers, cation exchange resins or polymers, neutral
exchange resins or polymers, immobilized polymyxin B, immobilized
monoclonal antibodies, immobilized inflammatory mediator receptors,
immobilized specific antagonists, cellulose, cellulose derivatives,
synthetic materials, polysulfone, polyacrylonitrile,
polymethylmethacrylate, polyvinyl-alcohol, polyamide,
polycarbonate, polystyrene-derivative fibers, and any combination
thereof. Specific examples of absorbent materials that have been
used in animal and clinical studies for non-specific binding of
inflammatory mediators, for example, include, but are not limited
to, polystyrene-divinylbenzene copolymer beads with biocompatible
polyvinylpyrrolidone coating (CYTOSORB, MedaSorb Corporation, NJ,
USA) and 2-methacryloyloxyethyl phosphorylcholine (MPCF-X; see,
e.g., Nakada, et al., Transfus. Apher. Sci. 35:253-264, 2006, which
is incorporated herein by reference.
[0183] In an aspect, the one or more binding agents can include one
or more biomolecules that non-specifically bind immunoglobulins,
including, but not limited to, anti-immunoglobulin antibodies,
Protein A and Protein G. In an aspect, the binding agent can be
avidin or streptavidin for binding biotinylated target components.
Alternatively, the binding agent can be biotin for binding avidin
or streptavidin labeled target components.
[0184] The one or more binding agents can be configured to
selectively bind one or more labeled target component. A selective
binding agent of one or more target cells can include, but is not
limited to, antibodies, antibody fragments, peptides,
oligonucleotides, DNA, RNA, aptamers, protein nucleic acids
proteins, receptors, receptor ligands, lectins, viruses, enzymes,
receptors, bacteria, cells, cell fragments, inorganic molecules,
organic molecules, an artificial binding substrate formed by
molecular imprinting, or other examples of biomolecules and/or
substrates that bind cells. The reactive components can include one
or more of an adhesion molecule, a binding mimetic, a polymer, a
lectin, integrin, or selectin.
[0185] The one or more binding agents can include one or more
antibodies that bind one or more labeled target components.
Antibodies, or fragments thereof, for use as one or more binding
agents labeled target components can include, but are not limited
to, monoclonal antibodies, polyclonal antibodies, Fab fragments of
monoclonal antibodies, Fab fragments of polyclonal antibodies,
Fab.sub.2 fragments of monoclonal antibodies, and Fab.sub.2
fragments of polyclonal antibodies, chimeric antibodies, non-human
antibodies, fully human antibodies, among others. Single chain or
multiple chain antigen-recognition sites can be used. Multiple
chain antigen-recognition sites can be fused or unfused. Antibodies
or fragments thereof can be generated using standard methods. See,
e.g., Harlow & Lane, Antibodies: A Laboratory Manual, Cold
Spring Harbor Laboratory Press; 1.sup.st edition 1988, which is
incorporated herein by reference. Alternatively, an antibody or
fragment thereof directed against one or more target component can
be generated, for example, using phage display technology. See,
e.g., Kupper, et al. BMC Biotechnology 5:4, 2005, which is
incorporated herein by reference. An antibody, a fragment thereof,
or an artificial antibody, e.g., Affibody.RTM. artificial
antibodies (Affibody AB, Bromma, Sweden) can be prepared using in
silico design (Knappik et al., J. Mol. Biol. 296: 57-86, 2000,
which is incorporated herein by reference. In an aspect, antibodies
directed against one or more target component may be available from
a commercial source (from, e.g., Novus Biological, Littleton,
Colo.; Sigma-Aldrich, St. Louis, Mo.; United States Biological,
Swampscott, Mass.).
[0186] The one or more binding agents can include one or more
aptamers that bind one or more labeled target components. The
aptamer can be an oligonucleotide RNA- or DNA-based aptamer.
Aptamers are artificial oligonucleotides (DNA or RNA) which can
bind to a wide variety of entities (e.g., metal ions, small organic
molecules, proteins, and cells) with high selectivity, specificity,
and affinity. Aptamers can be isolated from a large library of
10.sup.14 to 10.sup.15 random oligonucleotide sequences using an
iterative in vitro selection procedure often termed "systematic
evolution of ligands by exponential enrichment" (SELEX). See, e.g.,
Cao, et al., Current Proteomics 2:31-40, 2005; Proske, et al.,
Appl. Microbiol. Biotechnol. 69:367-374, 2005; Jayasena Clin. Chem.
45:1628-1650, 1999, each of which is incorporated herein by
reference.
[0187] In an aspect, the one or more binding agents can include one
or more aptamers that are peptide based aptamers. Peptide aptamers
are artificial proteins in which inserted peptides are expressed as
part of the primary sequence of a structurally stable protein. See,
e.g., Crawford, et al., Brief Funct. Genomic Proteomic 2:72-79,
2003, which is incorporated herein by reference.
[0188] The one or more binging agents can include one or more
peptide receptor ligands that bind receptors associated with one or
more target cells. Examples of peptide receptor ligands have been
described herein and generally include, but are not limited to,
neuropeptides, cytokines, chemokines, growth factors, and other
peptide hormones including atrial natriuretic factor, insulin,
glucagon, angiotensin, prolactin, oxyocin, and others. In an
aspect, the one or more binding agents can include one or more
novel peptides. Novel peptides that bind selective targets can be
generated using phage display methodologies. See, e.g., Spear, et
al., Cancer Gene Ther. 8:506-511, 2001, which is incorporated
herein by reference.
[0189] The one or more binding agents can include one or more
artificial binding substrates for one or more labeled target
component formed by the process of molecular imprinting. In the
process of molecular imprinting, a template is combined with
functional monomers which upon cross-linking form a polymer matrix
that surrounds the template, the process and materials of which
have been described herein. See, e.g., Alexander, et al., J. Mol.
Recognit. 19:106-180, 2006; U.S. Pat. No. 7,319,038; Byrne et al.,
Advanced Drug Delivery Reviews, 54: 149-161, 2002; U.S. Pat. Nos.
6,255,461; 5,804,563; 6,797,522; 6,670,427; and 5,831,012; U.S.
Patent Application 20040018508; Ye & Haupt, Anal Bioanal Chem.
378: 1887-1897, 2004; Peppas & Huang, Pharm Res. 19: 578-587
2002, each of which is incorporated herein by reference.
[0190] Reactive components can include denaturing agents that
modulate a physiological effect of the one or more target
components. The device including one or more reactive chemical
components can include one or more denaturing agents. Denaturing
includes changing one or more inherent characteristics or
properties of a cell, including cell constituents, protein, lipid,
or nucleic acid, to thereby negatively affect the cell and the
cellular components. Denaturing can include reducing a cell,
protein, lipid, or nucleic acid constituents to smaller components.
Denaturing can further include destroying cells, including cell
constituents, protein, lipid, or nucleic acid. Modulating a
physiological effect of the one or more target components can occur
by the process of denaturation in which the secondary, tertiary or
quaternary structure of the one or more target components can be
altered by denaturing agents. Examples of denaturing agents
include, but are not limited to, acids such as acetic acid,
trichloroacetic acid (TCA), sulfosalicyclic acid, picric acid;
solvents such as methanol, ethanol, and acetone; cross-linking
agents such as formaldehyde and gluteraldehyde; chaotropic agents
such as urea, guanidinium chloride, and lithium perchlorate; and
disulfide bond reducers such as 2-mercaptoethanol, dithithreitol,
TCEP, anionic detergents, or cationic detergents. In an aspect,
acids can be used to denature a protein molecule by exposing the
protein molecule to a pH below its isoelectric point. Under these
conditions, the protein molecule will lose its negative charge and
retain only positive charges. The like positive charges can repel
one another and in areas of large charge density, the
intramolecular repulsion can be sufficient enough to cause
unfolding of the protein. The one or more denaturing agents can be
incorporated into or released within one or more second reservoirs
of the device. Alternatively, the one or more denaturing agents can
be released by the device as diffusible agents into the blood or
lymph.
[0191] Reactive components can include degradative agents that
modulate a physiological effect of the one or more target
components. Modulating a physiological effect of the one or more
target components can occur by the one or more degradative agents
that act by breaking peptide bonds within the primary amino acid
sequence of the one or more target components. The one or more
degradative agents can include any of a number of agents designed
to cleave one or more peptide bonds of the primary amino acid
sequence of one or more target components. The one or more
degradative agents can include any of a number of agents designed
to cleave nucleic acids, lipids, or proteins associated with cells
or particles of the target component. Examples of degradative
agents, include, but are not limited to, proteases, strong acids,
strong bases, free radicals, natural or synthetic proteasomes, or
photoactivatable agents. The one or more degradative agents can be
incorporated into or released within one or more second reservoirs
of the device. Alternatively, the one or more degradative agents
can be released by the device as diffusible agents into the
blood.
[0192] The device including one or more degradative agents can
include one or more proteases. Examples of proteases include, but
are not limited to, serine proteases, e.g., as trypsin,
chymotrypsin, elastase, dipeptidyl peptidase IV, and subtilisin;
cysteine proteases, e.g., papain, cathepsins, caspases, calpains;
aspartic acid proteases, e.g., pepsin, renin, and HIV-proteases;
metalloproteases, e.g. carboxypeptidases, aminopeptidases, and
matrix metalloproteases, e.g. MMP1 through MMP28. The one or more
proteases can be free in solution. Alternatively, the one or more
proteases can be bound to a substrate. In an aspect, trypsin can be
bound to glass beads. See, e.g., Lee, et al., J. Dairy Sci.,
58:473-476, 1974, which is incorporated herein by reference.
Alternatively, trypsin and other proteases can be bound to an
agarose matrix. Sources of immobilized proteases including trypsin
and pepsin are available from commercial sources (Pierce Chemicals,
Rockford, Ill.; Applied Biosystems, Foster City, Calif.).
[0193] The device including one or more degradative agents can
include a natural or synthetic complex of proteases. In an aspect,
the one or more target components can be subject to degradation
using proteasomes. A proteasome is a naturally occurring large
protein complex that contains multiple subunits. The complex
includes several protease activities, for example,
chymotrypsin-like activity, trypsin-like activity, glutamic acid
protease activity, and threonine protease activity. Proteasome
complexes can be purified from fractionated cells using
ultracentrifugation through a 10-40% glycerol gradient. See, e.g.,
Pervan, et al., Mol. Cancer Res. 3:381-390, 2005, which is
incorporated herein by reference. Proteasomes can be isolated using
a commercially available isolation kit. (Proteasome Isolation Kit,
Human 539176-1KIT, Calbiochem (EMD Chemicals, Inc.; Gibbstown,
N.J.).
[0194] The device including one or more degradative agents can
include an agent that selectively targets one or more target
component for degradation. In an aspect, the one or more target
components can be covalently tagged with ubiquitin for selective
destruction by proteasomes. Ubiquitin is a small and highly
conserved protein. An isopeptide bond links the terminal carboxyl
of ubiquitin to the s-amino group of a lysine residue of a protein
targeted for degradation. The joining of ubiquitin to the targeted
protein is ATP-dependent. Three enzymes are involved, designated
E1, E2 and E3. Initially, the terminal carboxyl group of ubiquitin
is joined in an ATP-dependent thioester bond to a cysteine residue
on ubiquitin-activating enzyme (E1). The ubiquitin is then
transferred to a sulfhydryl group on a ubiquitin-conjugating enzyme
(E2). A ubiquitin-protein ligase (E3) then promotes transfer of
ubiquitin from E2 to the .epsilon.-amino group of a lysine residue
of a protein recognized by that E3, forming an isopeptide bond.
There are distinct ubiquitin ligases with differing substrate
specificity. In addition, some proteins have specific sequences
termed a "destruction box" that is a domain recognized by a
corresponding ubiquitin ligase. In general, E1, E2, and E3 can be
isolated from natural sources or generated using standard molecular
biology techniques and used to ubiquinate proteins in vitro. See,
e.g., Chen, et al., EMBO Rep. 2: 933-938, 2001, which is
incorporated herein by reference. In an aspect, the E2 ligase can
be genetically engineered in such a manner as to recognize a
specific substrate. See, e.g., Colas, et al., PNAS 97:13720-13725,
2000, which is incorporated herein by reference. The device
including the one or more degradative agents can include one or
more genetically engineered E2 ligase enzymes capable of adding
ubiquitin to and facilitating degradation of the one or more target
components in the blood of the vertebrate subject.
[0195] In an aspect, the ubiquitin can be indirectly associated
with the one or more target components. In an aspect, the ubiquitin
can be linked to an antibody or aptamer structural component of the
label. Binding of the ubiquitin-labeled antibody or aptamer to one
or more target components can mark the protein conjugate for
degradation by proteasomes.
[0196] The device including one or more degradative agents can
include a strong acid. Acid hydrolysis can result in degradation of
the one or more target components. In an aspect, strong acids such
as hydrochloric acid or sulfuric acid can be used to break the
carbon-nitrogen peptide bond. Degradation of one or more target
components by acid hydrolysis can be optionally performed in
combination with elevated temperature, a nitrogen atmosphere and/or
microwave energy.
[0197] The device including one or more degradative agents can
include one or more free radical reactive oxygen species. Examples
of reactive oxygen species include, but are not limited to, singlet
molecular oxygen, superoxide ion, hydrogen peroxide, hypochlorite
ion, hydroxyl radical. Reactive oxygen species can react directly
with proteins, targeting peptide bonds or amino acid side chains.
See, e.g., Davies, Biochem. Biophys. Res. Commun. 305:761-770,
2003, which is incorporated herein by reference. A number of the
reactions mediated by reactive oxygen species lead to introduction
of carbonyl groups into the protein which in turn can result in
inactivation of the protein by cleavage of the peptide bound to
yield lower-molecular weight products, cross-linking of proteins to
yield higher-molecular weight products, or loss of catalytic
function or structural function by distorting secondary and
tertiary structure, or combination thereof. Reactive oxygen species
can induce a amidation, diamide, glutamate oxidation and/or proline
oxidation which can lead to cleavage of peptide bonds. Reactive
oxygen species can be formed by the interaction of biological
molecules with components including, but not limited to, ionizing
radiation, as a byproduct of cellular respiration, and dedicated
enzymes such as NADPH oxidase and myeloperoxidase.
[0198] In an aspect, the device including one or more degradative
agents can include reactive oxygen species that are singlet oxygen
species. Singlet oxygen can cause damage to both the side-chains
and backbone of amino acids, peptides, and proteins. See, e.g.,
Davies, Biochem. Biophys. Res. Commun. 305:761-770, 2003, which is
incorporated herein by reference. Singlet oxygen species can react
with tryptophan, tyrosine, histidine, methionine and/or cysteine
and cystine residues within a polypeptide and can cause increased
susceptibility to proteolytic enzymes, an increased
extent/susceptibility to unfolding, changes in conformation, an
increase in hydrophobicity, and changes in binding of co-factor and
metal ions. In particular, the interaction of tyrosine with singlet
oxygen species can lead to fragmentation or cleavage of the
polypeptide. See, e.g., Davies, Biochem. Biophys. Res. Commun. 305:
761-770, 2003, which is incorporated herein by reference.
[0199] The device including one or more degradative agents can
include one or more singlet oxygen species generated by a
photosensitizer, a chemical which upon exposure to a given
wavelength of light emits singlet oxygen species. Examples of
photosensitizers include, but are not limited to, porphyrin
derivatives such as porfimer sodium, which is excited by red light
at 630 nm; chlorins and bacteriochlorins such as bonellin (maximum
absorbance 625 nm), mono-L-aspartyl chlorine e6 (max abs 654),
m-tetrahydroxyphenyl chlorine (mTHPC, max abs 652 nm), and tin
etiopurpurin (SnET2, maximum absorbance 660 nm); benzoporphyrin
derivatives such as veteroporfin (also labeled BPD-MA, maximum
absorbance 690 nm), 5-aminolaevulinic acid (ALA, porphoryin
precursor to PpIX (maximum absorbance 635 nm)); texaphyrins such as
lutetium texaphyrin (Lu-Tex, maximum absorbance 732),
Phthalocyanines and naphthalocyanines (maximum absorbance 670-780
nm); and cationic photosensitizers such as rhodamine 123 and
methylene blue. See, e.g., Prasad (2003) Introduction to
Biophotonics, John Wiley & Sons, Inc. Hoboken, N.J. Tunable
quantum dots (QDs), especially those absorbing in the wavelength
range of 600 to 800 nm, also emit singlet oxygen species in
response to light and can be useful as photosensitizers. See, e.g.,
Samia, et al. (2006) Photochem. Photobiol. 82:617-625, which is
incorporated herein by reference.
[0200] Reactive components can include target component-disrupting
agents that modulate a physiological effect of the one or more
target components. The device including one or more reactive
components can include one or more target component- or target
cell-disrupting agents. Examples of target component- or target
cell-disrupting agents include, but are not limited to, alcohols
and other organic solvents such as methanol, ethanol, isopropanol,
and acetone; cross-linking aldehydes such as formaldehyde and
gluteraldehyde; oxidizing agents such as sodium hypochlorite,
calcium hypochlorite, chloramine, chlorine dioxide, hydrogen
peroxide, iodine, ozone, acidic electrolyzed water, peracetic acid,
performic acid, potassium permanganate, potassium
peroxymonosulfate; acids such as acetic acid, trichloroacetic acid
(TCA), sulfosalicyclic acid, picric acid; phenolics such as phenol,
O-phenylphenol, chloroxylenol, hexachlorophene, thymol; chaotropic
agents such as urea, guanidinium chloride, and lithium perchlorate;
and disulfide bond reducers such as 2-mercaptoethanol,
dithiothreitol; and quaternary ammonium compounds. For example,
organic solvents such as methanol, ethanol or acetone can disrupt a
target component or target cell by solubilizing the lipids in the
plasma membrane and allowing the soluble contents of the cell to be
released. In an aspect, the one or more target component- or target
cell-disrupting agents are incorporated into or released within one
or more second reservoirs of the device. In an aspect, the one or
more target component- or target cell-disrupting agents are
released by the device as diffusible agents into the blood.
[0201] Modulating a physiological effect of the one or more target
component or target cells can occur by the one or more target
component- or target cell-disrupting agents breaking peptide bonds
within the primary amino acid sequence of proteins and peptides
associated with one or more target components or target cells. In
an aspect, the device including one or more target component- or
target cell-disrupting agents can include one or more proteases.
Examples of proteases include, but are not limited to, serine
proteases, e.g., as trypsin, chymotrypsin, elastase, dipeptidyl
peptidase IV, and subtilisin; cysteine proteases, e.g., papain,
cathepsins, caspases, calpains; aspartic acid proteases, e.g.,
pepsin, renin, and HIV-proteases; metalloproteases, e.g.
carboxypeptidases, aminopeptidases, and matrix metalloproteases,
e.g. MMP1 through MMP28. In an aspect, the one or more proteases
are free in solution. In an aspect, the one or more proteases are
bound to a substrate.
[0202] The device including one or more target component- or target
cell-disrupting agents can include one or more free radical
reactive oxygen species. Examples of reactive oxygen species
include, but are not limited to, singlet molecular oxygen,
superoxide ion, hydrogen peroxide, hypochlorite ion, hydroxyl
radical. Reactive oxygen species can react directly with proteins
associated with the one or more target cells, targeting peptide
bonds or amino acid side chains. In an aspect, the device including
one or more target component- or target cell-disrupting agents can
include reactive oxygen species that are singlet oxygen species.
Singlet oxygen can cause damage to both the side-chains and
backbone of amino acids, peptides, and proteins. See, e.g., Davies,
Biochem. Biophys. Res. Commun. 305: 761-770, 2003, which is
incorporated herein by reference. The target component- or target
cell-disrupting agents can include one or more singlet oxygen
species generated by a photosensitizer. Examples of
photosensitizers various classes of photosensitizers have been
described herein. A number of cell types including cancer cells and
bacterial pathogens are at least partially inactivated in response
to treatment with photosensitizers such as phthalocyanines,
phenothiazines, and porphyrins. See, e.g., Miller, et al., Toxicol.
Appl. Pharmacol. 224:290-299, 2007; Jori, et al., Lasers Surg. Med.
38:468-481, 2006; Keefe, et al., Lasers Surg. Med. 31:289-293,
2002, each of which is incorporated herein by reference.
[0203] Modulating a physiological effect of the one or more target
components. The device can include one or more reactive components
that can modulate a physiological effect of the one or more target
components in the blood of a vertebrate subject. The one or more
modulators are reactive components that can be incorporated into or
released within one or more second reservoirs of the device.
Alternatively, the one or more modulators can be released by the
device as diffusible agents into the blood. A modulator can alter,
modify, reduce or eliminate the activity of one or more target
components by preventing the binding of one or more target
components to their respective cognates. Alternatively, a modulator
can alter, modify, reduce or eliminate the activity of one or more
target components by inhibiting the enzymatic activity, e.g.,
phosphorylation activity, of the one or more target components.
Alternatively, the one or more modulators can indirectly alter,
modify or eliminate the activity of one or more target components
by attenuating the gene expression of one or more target
components. In an aspect, the one or more modulators can indirectly
alter or eliminate the activity of one or more target components by
increasing the expression of endogenous antagonists of the one or
more target components.
[0204] In general, the one or more modulator can be a protein, a
peptide, a small molecule, an aptamer, or an inhibitory RNA, DNA,
or nucleic acid. Modulators are contemplated that either directly
or indirectly antagonize the activity of one or more target
components and/or attenuate expression of one or more target
components.
[0205] In an aspect, the one or more modulator can be designed to
block the activity or binding properties of one or more target
components that is an inflammatory mediator. Examples of modulators
of inflammatory mediator activity and binding include, but are not
limited to, antibodies, e.g., infliximab, adalimumab, basiliximab
efalizumab; soluble receptors, e.g., etanercept, abatacept,
alefacept; corticosteroids, e.g., hydrocortisone, cortisone,
prednisone, prednisolone, methylprednisolone, meprednisone,
triamcinolone, paramethsone, fluprednisolone, betametasone, and
dexamethasone; nonsteroidal anti-inflammatory drugs (NSAIDS), e.g.,
selective cycloxygenase (COX) inhibitors exemplified by celecoxib,
etoricoxib, meloxicam, and valdecoxib and non-selective COX
inhibitors exemplified by diclofenac, difluisal, etodolac,
fenoprofen, fluripofen, ibuprofen, indomethacin, ketoprofen,
ketorolac, nabumetone, naproxen, oxaprozin, piroxicam, sulindac,
tenoxica, tiaprofen, tolmetin, azapropazone, and carprofen; and,
e.g., methotrexate, azathioprine, pennicillamine,
hydroxychloroquine, chloroquine, cyclophosphamide, cyclosporine,
mycophenolate mofetil, gold, and sulfasalazine.
[0206] Reactive chemical components can include cytotoxic,
cytostatic, apoptotic, and/or chemotherapeutic agents that modulate
a physiological effect of the one or more target cell components.
The device including one or more reactive chemical component,
reactive biologic components, or reactive physical components can
include one or more of a cytotoxic, a cytostatic, an apoptotic,
and/or a chemotherapeutic agent. Reactive chemical components,
reactive biologic components, or reactive physical components that
are cytotoxic, cytostatic, apoptotic, and/or chemotherapeutic
agents are contemplated that either directly or indirectly
inactivate or kill one or more target cells. Examples of cytotoxic,
cytostatic, apoptotic, and/or chemotherapeutic agents include, but
are not limited to, vinca alkaloids (e.g., vinblastine,
vincristine, vinflunine, vindesine, vinorelbine); taxanes (e.g.,
docetaxel, larotaxel, ortataxel, paclitaxel, tesetaxel);
epothilones (e.g., ixabepilone); dihydrofolate reductase inhibitors
(e.g., aminopterin, methotrexate, pemetrexed); thymidylate synthase
inhibitors (e.g., raltitrexed); adenosine deaminase inhibitor
(e.g., pentostatin); halogenated/ribonucleotide reductase
inhibitors (e.g., cladribine, clofarabine, fludarabine); thiopurine
(e.g., thioguanine, mercaptopurine); thymidylate synthase
inhibitors (e.g., fluorouracil, capecitabine, tegafur, carmofur,
floxuridine); DNA polymerase inhibitors (e.g., cytarabine);
ribonucleotide reductase inhibitor (e.g., gemcitabine,
hydroxyurea); hypomethylating agent (e.g., azacitidine,
decitabine); camptotheca (e.g., camptothecin, topotecan,
irinotecan, rubitecan, belotecan); podophyllum (e.g., etoposie,
teniposide); anthracyclines (e.g., aclarubicin, daunorubicin,
doxorubicin, epirubicin, idarubicin, amrubicin, pirarubicin,
valrubicin, zorubicin); anthracenediones (e.g., mitoxantrone,
pixantrone); nitrogen mustards (e.g., mechlorethamine,
cyclophosphamide, chlorambucil, bendamustine, uramustine,
estramustine); nitrosureas (e.g., carmustine, lomustine,
fotemustine, nimustine, ranimustine, streptozocin); aziridines
(e.g., carboquone, thioTEPA, triziquone, triethylenemelamine);
platinum (e.g., carboplatin, cisplatin, nedaplatin, oxaliplatin,
triplatin, tetranitrate, satraplatin); hydrazines (e.g.,
procarbazine); triazenes (e.g., dacarbazine, temozolomide,
altretamine, mitobronitol); streptomyces (actinomycin, bleomycin,
mitomycin, plicamycin); aminolevulinic acid/methyl aminolevulinate;
efaproxiral; porphyrin derivatives (porfimer sodium, talaporfin,
temoporfin, verteporfin); farnesyltransferase inhibitors,
cyclin-dependent kinase inhibitors, proteasome inhibitors,
phosphodiesterase inhibitors, IMP dehydrogenase inhibitors,
lipooxygenase inhibitors, PARP inhibitors, endothelin receptor
antagonists (e.g., atrasentan); retinoid X receptor (e.g.,
bexarotine); sex steroid (e.g., testolactone); amsacrine,
trabectedin, alitretinoin, tretinoin, arsenic trioxide, celecoxib,
demecolcine, elesclomol, elsamitrucin, etoglucid, lonidamine,
lucanthone, mitoguazone, mitotane, oblimersen, temsirolimus,
vorinostat. The cytotoxic agent can be a biological agent, e.g., a
peptide, a protein, an enzyme, a receptor and/or an antibody.
Examples of biological agents currently used to treat cancer
include, but are not limited to, cytokines such as
interferon-.alpha., interferon-.gamma., and interleukin-2, an
enzyme such as asparaginase, and monoclonal antibodies such as
alemtuzumab, bevacizumab, cetuximab, gemtuzumab, rituximab, and
trastuzumab.
[0207] The device including one or more reactive chemical
components, reactive biologic components, or reactive physical
components that include one or more cytotoxic, cytostatic,
apoptotic, and/or chemotherapeutic agents can include one or more
of an antibacterial drug. Examples of antibacterial drugs include,
but are not limited to, beta-lactam compounds (e.g., penicillin,
methicillin, nafcillin, oxacillin, cloxacillin, dicloxacilin,
ampicillin, ticarcillin, amoxicillin, carbenicillin, and
piperacillin); cephalosporins and cephamycins (e.g., cefadroxil,
cefazolin, cephalexin, cephalothin, cephapirin, cephradine,
cefaclor, cefamandole, cefonicid, cefuroxime, cefprozil,
loracarbef, ceforanide, cefoxitin, cefmetazole, cefotetan,
cefoperazone, cefotaxime, ceftazidine, ceftizoxine, ceftriaxone,
cefixime, cefpodoxime, proxetil, cefdinir, cefditoren, pivoxil,
ceftibuten, moxalactam, and cefepime); other beta-lactam drugs
(e.g., aztreonam, clavulanic acid, sulbactam, tazobactam,
ertapenem, imipenem, and meropenem); other cell wall membrane
active agents (e.g., vancomycin, teicoplanin, daptomycin,
fosfomycin, bacitracin, and cycloserine); tetracyclines (e.g.,
tetracycline, chlortetracycline, oxytetracycline, demeclocycline,
methacycline, doxycycline, minocycline, and tigecycline);
macrolides (e.g., erythromycin, clarithromycin, azithromycin, and
telithromycin); aminoglycosides (e.g., streptomycin, neomycin,
kanamycin, amikacin, gentamicin, tobramycin, sisomicin, and
netilmicin); sulfonamides (e.g., sulfacytine, sulfisoxazole,
silfamethizole, sulfadiazine, sulfamethoxazole, sulfapyridine, and
sulfadoxine); fluoroquinolones (e.g., ciprofloxacin, gatifloxacin,
gemifloxacin; levofloxacin, lomefloxacin, moxifloxacin,
norfloxacin, and ofloxacin); antimycobacteria drugs (e.g.,
isoniazid, rifampin, rifabutin, rifapentine, pyrazinamide,
ethambutol, ethionamide, capreomycin, clofazimine, and dapsone);
and miscellaneous antimicrobials (e.g., colistimethate sodium,
methenamine hippurate, methenamine mandelate, metronidazole,
mupirocin, nitrofurantoin, polymyxin B, clindamycin,
choramphenicol, quinupristin-dalfopristin, linezolid,
spectrinomycin, trimethoprim, pyrimethamine, and
trimethoprim-sulfamethoxazole).
[0208] The device including one or more reactive chemical
components, reactive biologic components, or reactive physical
components that include one or more cytotoxic, cytostatic,
apoptotic, and/or chemotherapeutic agents can include one or more
of an antifungal agent. Examples of antifungal agents include, but
are not limited to, anidulafungin, amphotericin B, butaconazole,
butenafine, caspofungin, clotrimazole, econazole, fluconazole,
flucytosine griseofulvin, itraconazole, ketoconazole, miconazole,
micafungin, naftifine, natamycin, nystatin, oxiconazole,
sulconazole, terbinafine, terconazole, tioconazole, tolnaftate,
and/or voriconazole.
[0209] The device including one or more reactive chemical
components, reactive biologic components, or reactive physical
components that include one or more cytotoxic, cytostatic,
apoptotic, and/or chemotherapeutic agents can include one or more
of an anti-parasite agent. Examples of anti-parasite agents
include, but are not limited to, antimalaria drugs (e.g.,
chloroquine, amodiaquine, quinine, quinidine, mefloquine,
primaquine, sulfadoxine-pyrimethamine, atovaquone-proguanil,
chlorproguanil-dapsone, proguanil, doxycycline, halofantrine,
lumefantrine, and artemisinins); treatments for amebiasis (e.g.,
metronidazole, iodoquinol, paromomycin, diloxanide furoate,
pentamidine, sodium stibogluconate, emetine, and dehydroemetine);
and other anti-parasite agents (e.g., pentamidine, nitazoxanide,
suramin, melarsoprol, eflornithine, nifurtimox, clindamycin,
albendazole, and tinidazole).
[0210] The device including one or more reactive chemical
components, reactive biologic components, or reactive physical
components that include one or more cytotoxic, cytostatic,
apoptotic, and/or chemotherapeutic agents can include one or more
of an antiviral agent. Examples of antiviral agents include, but
are not limited to, nucleoside analogs used to treat herpes simplex
virus (HSV) and varicella-zoster virus (VZV) (e.g., valacyclovir,
famciclovir, penciclovir, and trifluridine); nucleoside analogs
used to treat cytomegalovirus (CMV) (e.g., ganciclovir,
valganciclovir, and cidofovir); nucleoside and nonnucleoside
reverse transcriptase inhibitors used to treat HIV (e.g., abacavir,
didanosine, emtricitabine, lamivudine, stavudine, tenofovir,
zalcitabine, zidovudine, delavirdine, efavirenz, and nevirapine);
protease inhibitors used to treat HIV (e.g., atazanavir, darunavir,
fosamprenavir, indinavir, lopinavir, nelfinavir, ritonavir,
saquinavir, and tipranavir); and drugs used to treat hepatitis
(e.g., interferon alfa, adefovir dipivoxil, entecavir, and
ribavirin).
[0211] Two or More Reactive Components Combined to Modulate a
Physiological Effect of the One or More Target Components. The
device can include two or more reactive components that have been
combined to modulate a physiological effect of the one or more
target components. The two or more combined reactive components can
be one or more binding agent combined with one or more of a
denaturing agent, a degradative agent, a cell-disrupting agent, a
modulator, an apoptotic agent, a cytotoxic agent, cytostatic agent,
a chemotherapeutic agent, an antibody toxin or a combination
thereof.
[0212] In an aspect, the device can include the two or more
reactive components incorporated into a single biomolecule. For
example, the first reactive component can be a binding agent, e.g.,
an antibody, that includes a second reactive component that is a
degradative activity. Certain antibodies are capable of cleaving
the amide bond of peptide bonds. See, e.g., Janda, et al., Science
241:1188-1191, 1988; Lacroix-Desmazes, et al., J. Immunol. 177:
1355-1363, 2006; Ponomarenko, et al., Proc. Natl. Acad. Sci. USA,
103: 281-286, 2006; and U.S. Pat. No. 6,387,674, each of which is
incorporated herein by reference. Alternatively, the first reactive
component can be a binding agent, e.g., an antibody, and can
include a second reactive component that is a reactive oxygen
species. The one or more target components can bind to one or more
binding agents that are catalytic antibodies capable of generating
the reactive oxygen species H.sub.2O.sub.2 in response to UV
radiation. See, e.g., Wentworth, et al., Science 293: 1806-1811,
2001; Wentworth, Science 296: 2247-2249, 2002; Wentworth, et al.,
Proc. Natl. Acad. Sci. USA, 97: 10930-10935, 2000, each of which is
incorporated herein by reference. One or more antibodies or other
binding agents can be generated for both binding and degradation of
one or more labels.
[0213] In an aspect, the first reactive component can be a binding
agent, e.g., an antibody, that can include a second reactive
component that is a cellular toxin. For example, the first reactive
component can be an antibody that binds a cellular target component
and the second reactive component can be a photosensitizer which is
activated upon exposure to electromagnetic energy. See, e.g.,
Serebrovskaya, et al., Proc. Natl. Acad. Sci. USA. 106:9221-9225,
2009, which is incorporated herein by reference. In another
example, the first reactive component can be an antibody directed
against a cancer cell or other cellular target component and the
second reactive component can be an auristatin, a cytotoxic
inhibitor of tubulin polymerization. See, e.g., Ma, et al., Clin.
Cancer Res. 12:2591-2596, 2006, which is incorporated herein by
reference.
[0214] In an aspect, the two or more reactive components of the
device can be incorporated into a single biomolecule and can
include a first component that is a binding agent, e.g., an
aptamer, and a second component that is a degradative agent, e.g.,
a protease. For example, one or more proteases can be conjugated or
chemically linked to one or more oligonucleotide-based aptamers.
The oligonucleotide-based aptamers are designed to bind to the
target component. Upon binding to the oligonucleotide-based
aptamers, the one or more target components are brought into
proximity to the one or more proteases resulting in proteolytic
degradation of the one or more target components. Proteases can be
linked to oligonucleotide-based aptamers using any of a number of
methods for conjugating a polypeptide to an oligonucleotide. In an
aspect, a polypeptide protease can be conjugated to an
oligonucleotide-based aptamer using a streptavidin-biotin bridge by
introducing a biotinylated oligonucleotide into the aptamer
sequence and linking it to a biotinylated protease through a
streptavidin bridge. Alternatively, the polypeptide protease can be
conjugated to the oligonucleotide-based aptamer using a
thiol-maleimide linkage in which a carbon with an attached thiol
group is placed on the aptamer and reacts with a maleimide group
added to the C terminus of the protease. See, e.g., Nitin, et al.,
Nucleic Acids Res. 32:e58, 2004, which is incorporated herein by
reference. A number of modified nucleotides are commercially
available for use in synthesizing oligonucleotide aptamers with
amines or other side chains for cross-linking (TriLink
Biotechnologies, San Diego, Calif.; Sigma Aldrich, St. Louis,
Mo.).
[0215] In an aspect, the first reactive component can be a binding
agent linked to a second reactive component and encapsulated in a
tunable vesicle. For example, the second reactive component, e.g.,
a denaturing and/or degradative agent, can be encapsulated in a
tunable hydrogel. The binding of one or more target components to
the label is illuminated by the energy source and leads to release
of the denaturing and/or degradative agent from the hydrogel. In an
aspect, target-responsive hydrogels can be generated in which the
contents of the hydrogel are selectively released in response to
binding a specific target. The hydrogel can incorporate one or more
binding agents that are antibodies. The hydrogel can release its
contents in response to an antibody-antigen interaction. See, e.g.,
Miyata, et al., Proc. Natl. Acad. Sci. USA 103: 1190-1193, 2006,
which is incorporated herein by reference. In an aspect, the
target-responsive hydrogel can incorporate one or more binding
agents that are oligonucleotide-based aptamers and release its
contents in response to an aptamer-ligand interaction. See Yang, et
al., J. Am. Chem. Soc. 130:6320-6321, 2008, which is incorporated
herein by reference. In the latter case, two or more distinct
aptamers configured to partially overlap during hybridization can
be copolymerized into a polyacrylamide hydrogel. At least one of
the two or more aptamers binds to a specific target, e.g., target
component. When the target component binds to the aptamer, the
number of nucleotide bases available for hybridization between the
overlapping aptamers is reduced, causing them to separate.
Separation of the overlapping aptamers allows the hydrogel to
dissolute and release its contents. A target responsive hydrogel
can be generated which incorporates aptamers that specifically
recognize one or more target components. The hydrogel itself can be
loaded with one or more proteases or other reactive components that
are configured to modulate a physiological effect of the target
components. The contents of the hydrogel are released upon binding
of the one or more target components to the aptamers associated
with the hydrogel. In an aspect, hydrogels can be used for
molecular imprinting. See, e.g., Byrne et al., "Molecular
imprinting within hydrogels," Advanced Drug Delivery Reviews, 54:
149-161, 2002, which is incorporated herein by reference.
Device Including Substrates for Reactive Components
[0216] The device can include one or more reactive components
including binding agents, denaturing agents, degradative agents,
modulators, or combinations thereof, wherein the one or more
reactive components can be free in solution within one or more
second reservoirs of the device. Alternatively, the one or more
reactive components can be immobilized on a solid substrate within
the one or more second reservoirs or within the lumen of the
device. The solid substrate can be a matrix, e.g., a bead or a
filter, added to one or more second reservoirs of the device.
Examples of applicable solid substrates include, but are not
limited to, beads, particles, membranes, semi-permeable membranes,
capillary, or microarrays. The solid substrate can be comprised of
an inorganic material, e.g., glass, alumina, silica, silicon,
zirconia, graphite, magnetite, semiconductors, or combinations
thereof. Alternatively, the solid substrate can be comprised of an
organic material, e.g., polysaccharides including agarose, dextran,
cellulose, chitosan, and polyacrylamide, polyacrylate, polystyrene,
polyvinyl alcohol, or combinations thereof. Alternatively, the one
or more specific binding agents or one or more reactive components
can be associated with a solid substrate that are cells, e.g.,
mammalian cells, enucleated erythrocytes, bacteria, or viral
particles or vesicles such as liposomes or other micellular
vesicles.
[0217] In an aspect, the one or more reactive components, either
free in solution or bound to a solid substrate, can be prevented
from leaving the one or more second reservoirs of the device either
due to size exclusion using a filter or mesh or due to physical
attachment to the device. In a detailed aspect, one or more labeled
target components present in the blood can bind to the one or more
reactive components and be sequestered for inactivation as the
blood passes through the device.
[0218] The one or more reactive components can be bound to the
solid substrate either directly or indirectly. For example, the one
or more reactive components can be coupled to the solid substrate
by covalent chemical bonds between particular functional groups on
the specific binding agent (e.g., primary amines, sulfhydryls,
carboxylic acids, hydroxyls, and aldehydes) and reactive groups on
the solid substrate. A variety of activating compounds and schemes
for directly bonding ligands to solid substrates are known. Some
examples include, but are not limited to, cyanogen bromide,
cyanuric chloride, epichlorohydrin, divinyl sulphone,
p-toluenesulphonyl chloride, 1,1'-carbonyldiimidazole, sodium
meta-periodate, 2-fluoro-1-methylpyridiniumtoluene-4-sulphonate,
glycidoxypropyl-trimethoxysilane and 2,2,2-trifluoroethanesulphonyl
chloride. For example, cyanogen bromide in base reacts with
hydroxyl (OH) groups on agarose solid substrate to form cyanate
esters or imidocarbonates. These groups readily react with primary
amines under mild conditions resulting in a covalent coupling of
the ligand to the agarose solid substrate. Reactive imidocarbonates
can also be formed on carbon nanotubes, for example, through
reactive carboxyl groups generated by treatment of the nanotubes
with oxidizing agents. See, e.g., Bianco, et al., in Nanomaterials
for Medical Diagnosis and Therapy. pp. 85-142. Nanotechnologies for
the Live Sciences Vol. 10 Edited by Challa S. S. R. Kumar,
WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, 2007, which is
incorporated herein by reference. Functionalization of silicon
chips with carboxyl groups can be subsequently used to immobilize
proteins in the presence of
N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC)
and N-hydroxysuccinimide ester (NHS). See, e.g., Hu, et al., Rapid
Commun. Mass Spectrom. 21:1277-1281, 2007, which is incorporated
herein by reference.
[0219] The one or more reactive components can have linking or
spacer groups bound to the C-terminus which when present can be
used to bind the specific binding agent to the solid substrate
indirectly. When present the linking group can be a polymer or a
monomer. A linking group can be a chain of from 1-10 amino acids.
Other examples of linking groups include, but are not limited to,
polyethylene glycol, polypropylene glycol, polyesters,
polypeptides, polyethers, polysaccharides, glycidoxyalkyl,
alkoxyalkyl, alkyl, glycidoxypropyl, ethyl, propyl, phenyl and
methacryl; and silicon containing linking groups such as
diethyl(triethoxysilylpropyl)malonate;
3-mercaptopropyltrimethoxysilane; 3-aminopropyltrimethoxysilane;
N-[(3-trimethoxysilyl)propyl]ethylenediaminetriacetic acid;
p-(chloromethyl)phenyltrimethoxysilane; vinyltriethoxysilane;
3-bromopropyltriethoxysilane; and
3-glycidoxypropyltrimethoxysilane.
[0220] In general, any of a number of homobifunctional,
heterofunctional, and/or photoreactive cross linking agents can be
used to conjugate one or more reactive components to an
appropriately derivatized substrate. Examples of homobifunctional
cross linkers include, but are not limited to, primary
amine/primary amine linkers such as BSOCES
((bis(2-[succinimidooxy-carbonyloxy]ethyl)sulfone), DMS (dimethyl
suberimidate), DMP (dimethyl pimelimidate), DMA (dimethyl
adipimidate), DSS (disuccinimidyl suberate), DST (disuccinimidyl
tartate), Sulfo DST (sulfodisuccinimidyl tartate), DSP
(dithiobis(succinimidyl propionate), DTSSP
(3,3'-dithiobis(succinimidyl propionate), EGS (ethylene glycol
bis(succinimidyl succinate)) and sulfhydryl/sulfhydryl linkers such
as DPDPB (1,4-di-(3'-[2' pyridyldithio]-propionamido) butane).
Examples of heterofunctional cross linkers include, but are not
limited to, primary amine/sulfhydryl linkers such as MBS
(m-maleimidobenzoyl-N-hydroxysuccinimide ester), Sulfo MBS
(m-maleimidobenzoyl-N-hydroxysulfosuccinimide), GMBS
(N-gamma-maleimidobutyryl-oxysuccinimide ester), Sulfo GMBS
(N-y-maleimidobutyryloxysulfosuccinimide ester),
EMCS(N-(epsilon-maleimidocaproyloxy)succinimide ester), Sulfo
EMCS(N-(epsilon-maleimidocaproyloxy)sulfo succinimide), SIAB
(N-succinimidyl(4-iodoacetyl)aminobenzoate), SMCC (succinimidyl
4-(N-maleimidomethyl)cyclohexane-1-carboxylate), SMPB (succinimidyl
4-(rho-maleimidophenyl)butyrate), Sulfo SIAB
(N-sulfosuccinimidyl(4-iodoacetyl)aminobenzoate), Sulfo SMCC
(sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate),
Sulfo SMPB (sulfosuccinimidyl 4-(rho-maleimidophenyl)butyrate), and
MAL-PEG-NHS (maleimide PEG N-hydroxysuccinimide ester);
sulfhydryl/hydroxyl linkers such as PMPI
(N-rho-maleimidophenyl)isocyanate; sulfhydryl/carbohydrate linkers
such as EMCH(N-(epsilon-maleimidocaproic acid) hydrazide); and
amine/carboxyl linkers such as EDC
(1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride).
[0221] In an aspect, the one or more reactive components can be
linked to a solid substrate through non-covalent interactions.
Examples of non-covalent interactions include, but are not limited
to, protein-protein interactions such as those between
avidin/streptavidin and biotin, protein A and immunoglobulins,
protein G and immunoglobulins, or secondary antibodies with primary
antibodies. For example, the one or more reactive components can be
modified with biotin using standard methods and bound to a solid
substrate derivatized with streptavidin. One or more reactive
components can be modified with streptavidin and bound to a solid
substrate derivatized with biotin. A single chain antibody can
incorporate streptavidin as part of a fusion protein to facilitate
attachment of the antibody to the solid substrate via a
biotin-streptavidin linkage. See, e.g., Koo, et al. Appl. Environ.
Microbiol. 64:2497-2502, 1999. Solid substrates such as beads or
other particulate substrates derivatized with protein A, protein G,
streptavidin, avidin, biotin, secondary antibodies are available
from commercial sources (from, e.g., Pierce-Thermo Scientific,
Rockford, Ill., Sigma-Aldrich, St. Louis, Mo.). In an aspect, the
one or more reactive components can bind to the solid substrate
through a non-covalent interaction and be cross-linked to the solid
substrate using a cross-linking agent.
[0222] In an aspect, the one or more reactive components can be
associated with cells as a solid substrate, e.g., mammalian cells,
enucleated erythrocytes, bacteria, or viral particles, or vesicles
such as liposomes or other micellular vesicles. Cells and vesicles
can be modified with one or more reactive components using many of
the same methods as provided herein. One or more reactive
components can be bound to cells or vesicles using one or more
homobifunctional or heterofunctional cross-linkers through primary
amines and carboxyl groups. Alternatively, cells can be modified
with one or more reactive components using a biotin-streptavidin
bridge. For example, one or more reactive components can be
biotinylated and linked to a non-specifically biotinylated cell
surface through a streptavidin bridge. An antibody, aptamer, or
receptor can be biotinylated using standard procedures. The surface
membrane proteins of a cell can be biotinylated using an amine
reactive biotinylation reagent such as, for example, EZ-Link
Sulfo-NHS-SS-Biotin (sulfosuccinimidyl
2-(biotinamido)-ethyl-1,3-dithiopropionate; Pierce-Thermo
Scientific, Rockford, Ill., USA; see, e.g., Jaiswal, et al. Nature
Biotech. 21:47-51, 2003; U.S. Pat. No. 6,946,127).
[0223] In an aspect, the one or more reactive components can be
associated with lipid or micellular vesicles. In an aspect, the one
or more reactive components can be antibodies attached to a
liposome. Antibodies can be added to liposomes using cross-linking
agents and protein A. See, e.g., Renneisen, et al., J. Bio. Chem.,
265:16337-16342, 1990, which is incorporated herein by reference.
The liposomes are formed from dry lipid in the presence of an
aqueous solution, e.g., a buffer of appropriate pH followed by
extrusion through a high pressure device fitted with a
polycarbonate filter with the desired pore size to form liposomes
of a specific size range. The liposomes are modified with
N-succinimidyl 3-(2-pyridyldithio)propionate-modified protein A.
The one or more antibodies are linked to the liposomes through
selective binding to the protein A. Alternatively, thiolated
antibodies can be covalently linked to liposomes prepared with
4-(p-maleimidophenyl)butyrylphosphatidyl-ethanolamine. See, e.g.,
Heath, et al., PNAS 80:1377-1381, 1983, which is incorporated
herein by reference.
[0224] In an aspect, the one or more reactive components can be
expressed on the surface of a cell. The one or more reactive
components can be naturally expressed on the surface of a cell,
such as a receptor of a specific inflammatory mediator on a
specific cell type. Alternatively, the one or more reactive
components can be expressed on the surface of a cell using genetic
manipulation. For example, cells can be genetically manipulated to
express a receptor that binds one or more structural elements of
the target component. In one example, cells can be genetically
manipulated to express one or more specific antibodies on the cell
surface. Methods have been provided for cell surface expression of
single chain Fv antibody fragments (scFv) fused to
membrane-associated proteins. See, e.g., Ho, et al., Proc. Natl.
Acad. Sci. USA 103:9637-9642, 2006; Francisco, et al., Proc. Natl.
Acad. Sci. USA 90:10444-10448, 1993; U.S. Pat. Appl. No.
2006/0083716, each of which is incorporated herein by reference. In
a detailed aspect, the cDNA sequence encoding all or part of an
antibody recognizing a structural element of the target component
is fused in an expression construct in frame with a
membrane-associated protein and expressed in an appropriate cell
type.
Device Including Controller in Communication with and Responsive to
a Sensor
[0225] The device can include at least one controller in
communication with the one or more sensors, and in communication
with the at least one controllable flow barrier to the at least one
lumen, wherein the controller is configured to control flow of the
one or more of blood or lymph through the at least one lumen. The
one or more sensors can transmit data to the controller regarding
the one or more signal responses associated with the one or more
labeled target components in the blood or lymph of the vertebrate
subject. The controller can be integrated into the device.
Alternatively, the controller can be a separate component of the
device that receives and transmits data and/or commands either with
or without wires. For example, an implanted device can send data
regarding the sensed signal responses of one or more target
components to an external controller through a wireless signal.
[0226] The controller can compare the input data regarding the
physiological effect of the one or more target components in the
blood or lymph of the vertebrate subject with stored data. The
controller itself can include the stored data. Alternatively, the
controller can have access to one or more remote databases that
include the stored data. The stored data can be data regarding the
normal levels of one or more target components in normal or healthy
subjects without a disease or condition. The stored data can be
data regarding normal parameters, e.g, size, granularity,
autofluorescence, scattering properties of one or more target
components in normal or healthy subjects. The stored data can
include data regarding the baseline level of one or more target
components in a vertebrate subject prior to the onset of a disease
or condition. The stored data can include data regarding the level
of one or more target components in a vertebrate subject at one or
more previous time points. The controller assesses the most
recently obtained input data with the stored data and is configured
to controllably initiate steps to release an amount of one or more
labels from the at least one first reservoir.
[0227] The device including the controller can also be in
communication with and configured to be informed by one or more
sensors. In response to input data, the controller can cause the
device to controllably divert all or part of the blood or lymph of
a vertebrate subject through the controllable flow barrier into at
least one lumen. Access to at least one lumen can be controlled by
a flow-modulating element or controllable flow barrier. A
flow-modulating element can be a gate, a valve, a louver, a
splitter or flow divider, a filter, a baffle, a channel
restriction, a retractable iris, or other structure that
controllably limits access of the blood flow to at least one lumen.
The controller can send a signal to the flow-modulating element
indicating whether or not all or part of the flow of blood should
be diverted into at least one lumen.
[0228] The device including the controller can also be in
communication with and configured to be informed by one or more
sensors to control access to or release of one or more reactive
components from one or more second reservoirs. In response to input
data, the controller can controllably initiate release or
activation of one or more reactive components designed to modulate
a physiological effect of the one or more target components, e.g.,
to inactivate, ablate, alter, arrest, disrupt, destroy, inactivate,
or ablate the one or more target components. The one or more
reactive components are controllably released or activated in the
one or more second reservoirs of the device. In an aspect, the
controller can release one or more reactive components to modulate
the activity of one or more target components. Alternatively, the
controller can send data regarding the levels of one or more target
components in the blood of the vertebrate subject to the subject,
to one or more third party individuals such as a physician or other
caregiver, to a computing device, or to a combination thereof. The
subject and/or caregiver or computing device can choose to initiate
steps to inactivate, ablate, alter, arrest, disrupt, destroy,
inactivate, or ablate the one or more target components by
releasing or activating one or more reactive components.
[0229] The controller can also include one or more algorithms that
provide computational models of a disease or condition. For
example, a computational model of a disease or condition can
include information regarding, for example, a variety of
interrelated signaling pathways involved in the disease process.
The computational model can inform decisions made by the
controller. Examples of computational models related to
inflammatory disease, cancer and pathogen infection have been
described. See, e.g., U.S. Pat. No. 7,415,359 B2; U.S. Patent
Applications 2007/0083333 A1, 2008/0201122 A1; Vodovotz, et al.,
Curr. Opin. Crit. Care. 10:383-390, 2004; Zenker, et al., PLoS
Comput. Biol. 3(11):e204, 2007; Li, et al., PLoS ONE 3(7):e2789,
2008; Vodovotz, et al., PLoS Comput. Biol. 4:e1000014, 2008; An,
Theoretical Biology Medical Modeling 5:11, 2008; Lee, et al., Proc.
Natl. Acad. Sci. USA. 104:13086-13091, 2007; Zhou, et al., HIV
Medicine. 6:216-223, 2005, each of which is incorporated herein by
reference.
Device for Controlling Levels of One or More Target Components to a
Target Level
[0230] A device is disclosed herein that includes a sensor
configured to detect one or more signal response associated with
one or more labeled target components in the blood or lymph of a
vertebrate subject. The device is configured to modulate a
physiological effect of the one or more target components to
control levels of the one or more target components to a target
value. The target value can be a desired concentration of one or
more target components in the blood or lymph, or the target value
can be a desired range of concentrations of one or more target
components in the blood or lymph. Alternatively, the target value
can be a desired ratio of concentrations of two or more target
components in the blood or lymph. The desired ratio can be
determined by a least squares fit of the concentrations of the two
or more target components. The target value of a target component
can be a desired concentration and/or concentration range and/or
ratio of concentrations that is a specific value or range of values
such as, for example, a value or range of values observed in a
normal subject. Alternatively, the target value of a target
component can be a desired concentration and/or concentration range
and/or ratio of concentrations that is at least 20%, at least 40%,
at least 60%, at least 80%, or at least 100% below or above the
current level of the target component in the blood or lymph of a
vertebrate subject.
[0231] The target value of one or more target components can be a
desired concentration and/or concentration range that is below that
observed in the blood or lymph of a vertebrate subject experiencing
a disease, disorder, or infection. For example, a number of target
components, e.g., inflammatory mediators, are elevated in the blood
of subjects diagnosed with systemic immune response syndrome (SIRS)
and sepsis. See, e.g., Ueda, et al., Am. J. Respir. Crit. Care Med.
160:132-136, 1999; Kurt, et al., Mediators Inflamm. 2007:31397,
2007; Kellum, et al., Arch. Intern. Med. 167:1655-1663, 2007; Wang,
et al., Crit. Care 12:R106, 2008; each of which is incorporated
herein by reference. As an example, the levels of TNF-.alpha.,
IL-6, and IL-8 in normal subjects is reported as less than 5 pg/ml,
less than 10 pg/ml, and less than 10 pg/ml, respectively. In
individuals with septic shock, the serum levels of TNF-.alpha.,
IL-6, and IL-8 are significantly elevated to mean values of
138+/-22 pg/ml, 27,255+/-7,895 pg/ml, and 2,491+/-673 pg/ml,
respectively. See Ueda, et al., Am. J. Respir. Crit. Care Med.
160:132-136, 1999; which is incorporated herein by reference.
[0232] The relative levels of one or more target components in the
blood or lymph of the vertebrate subject can be correlated with
prognosis and survival. For example, sepsis non-survivors have
proportionally higher levels of inflammatory mediators relative to
sepsis survivors and normal controls. In one study, high levels of
both IL-10 (mean of 45 pg/ml) and IL-6 (mean of 735 pg/ml) at
hospital admission were associated with increased mortality as
compared with low initial levels of IL-10 (mean of 7.4 pg/ml) and
IL-6 (mean of 15 pg/ml). In another example, elevated serum levels
of IL-6 were correlated with sepsis symptom scores and poor
outcome. These data suggest that modulating the levels of one or
more inflammatory mediators to a desired target value in the blood
can alter the course of the disease. See, e.g., Kellum, et al.,
Arch. Intern. Med. 167:1655-1663, 2007; Presterl, et al., Am. J.
Respir. Crit. Care Med. 156:825-832, 1997; each of which is
incorporated herein by reference.
[0233] The relative levels of one or more target components in the
blood of the vertebrate subject can be correlated with a chronic
disease. For example, subjects with rheumatoid arthritis have
increased levels of various inflammatory mediators relative to
normal subjects including IL-6 (15.8 pg/ml versus 4.0 pg/ml),
TNF-.alpha. (10 pg/ml versus 3.2 pg/ml), IL-1.beta. (129.8 pg/ml
versus 57.3 pg/ml), IL-8 (9.3 pg/ml versus 2.6 pg/ml), IL-10 (15.5
pg/ml versus 4.6 pg/ml), and IL-12 (20.2 pg/ml versus 6.2 pg/ml).
Psoriatic arthritis is also characterized by increased levels of
circulating inflammatory mediators with statistically significant
increases in the serum levels of various inflammatory mediators in
subjects diagnosed with psoriatic arthritis versus normal. See,
e.g., Nowlan, et al., Rheumatology 45:31-37, 2006; Mittal &
Joshi, J. Indian Rheumatol. Assoc. 10:59-60, 2002; Szodoray, et
al., Rheumatology 46:417-425, 2007, each of which is incorporated
herein by reference.
[0234] The target value of one or more cellular target components
can be a desired concentration or concentration range of target
cells that is below that observed in the blood or lymph of a
vertebrate subject experiencing a disease or condition. For
example, elevated levels of red blood cells are associated with
exposure to carbon monoxide, long-term lung disease, kidney
disease, some cancers, certain forms of heart disease, liver
disease. Elevated levels of platelets are associated with bleeding,
iron deficiency, some diseases like cancer, or bone marrow
problems. Elevated levels of neutrophils, eosinophils, and/or
lymphocytes are associated with infection, malignancy and
autoimmune diseases. The desired concentration or concentration
range can be the concentration or concentration range observed in a
normal individual. For example, the normal range of white blood
cells in men and nonpregnant women ranges from 4.5 to
11.times.10.sup.9 cells per liter while in pregnant women, the
white blood cell counts range from 5.9 to 25.7.times.10.sup.9 cells
per liter depending upon whether the vertebrate subject is in the
first, second or third trimester or postpartum. Similarly, normal
red blood cell counts range from 4.7 to 6.1.times.10.sup.12 in men,
4.2 to 5.4.times.10.sup.12 in women, 4.0 to 5.5.times.10.sup.12 in
children and 4.8 to 7.1.times.10.sup.12 in newborns. Normal
platelet counts range from 150 to 450.times.10.sup.9 for children
and 150 to 400.times.10.sup.9 for adults. See, e.g., Rea, WebMD.
Complete Blood Count (CBC) at
www.webmd.com/a-to-z-guides/complete-blood-count-cbc. Last updated
Sep. 12, 2008; accessed Oct. 5, 2009; incorporated herein by
reference.
[0235] The target value can be a percentage range of cells in the
blood. For example, of the total white blood cells in a normal
subject, neutrophils range from 50% to 62%, band neutrophils range
from 3% to 6%, lymphocytes range from 25% to 40%, monocytes range
from 3% to 7%, eosinophils range from 0% to 3%, and basophils range
from 0% to 1%.
[0236] The target value can be a desired ratio of concentrations of
two or more target components in the blood or lymph as determined
by a least squares fit of the concentration values of the two or
more target components. In this instance, the levels of one or more
target components can be altered to modulate the overall ratio of
two or more target components. For example, levels of neutrophils
relative to leukocytes is reportedly is correlated with
cardiovascular risk in that increased neutrophils and/or decreased
leukocytes are associated with diabetes, coronary artery disease,
unstable angina, and increased risk of myocardial infarction. See,
e.g., Horne, et al., J. Am. Coll. Cardiol. 45:1638-1643, 2005,
which is incorporated herein by reference.
[0237] In some pathological states such as cancer or infection, the
ideal target value of one or more target components can be zero. In
the instance where a target value of zero is not attainable, the
target value can be a value that reduces the symptoms and/or the
disease progression. In malaria infected individuals, for example,
the degree of parasitemia is correlated with the severity of the
disease. The number of parasites per microliter of blood is used to
assess parasitemia. For example, a vertebrate subject can just be
showing signs of symptoms at 100 parasites per microliter (0.002%
parasitemia), severe malaria at 100,000 to 250,000 parasites per
microliter (2-5% parasitemia), and near death at 500,000 parasites
per microliter (10% parasitemia). Reducing parasitemia can reduce
symptoms and disease severity.
[0238] Similarly, the target value of one or more target component,
e.g., a toxin or illicit drug, can be zero. In the instance where a
target value of zero is not attainable, the target value can be a
value that reduces toxicity. For example, elevated levels of lead
in the blood in adults can damage the nervous, hematologic,
reproductive, renal, cardiovascular, and gastrointestinal systems.
The majority of cases of lead poisoning are workplace related. The
U.S. Department of Health and Human Services recommends that blood
levels of lead among adults be reduced to <25 ug/dL. The highest
blood levels of lead acceptable by standards of the U.S.
Occupational Safety Health Administration is 40 ug/dL. The
geometric mean blood levels of lead of all adults in the U.S. is
<3 ug/dL. MMWR 58(14):365-369, 2009, which is incorporated
herein by reference.
PROPHETIC EXAMPLES
Example 1
Device Including Sensor for Detecting One or More Target
Circulating Tumor Cells and for Binding and Altering Circulating
Tumor Cells for Treatment of a Neoplastic Disease or Condition
[0239] An implantable device is described for treating a neoplastic
disease, e.g. metastatic breast cancer, associated with the
presence of circulating tumor cells in the peripheral blood of a
subject. The components of the implantable device are incorporated
into a stent inserted into a femoral artery of the subject. The
implantable device includes two reservoirs: a first reservoir
containing a flourescein-labeled aptamer that can bind a
circulating tumor cell in the peripheral blood of a subject and
emit a fluorescent signal in response to the binding; and a second
reservoir containing a chemotherapeutic agent, e.g., doxorubicin.
The implantable device further includes an optical sensor
consisting of a set of light emitting diodes (LEDs) emitting light
at a specific wavelength to elicit a signal response associated
with the fluorescently labeled circulating tumor cells, and a
metal-oxide-semiconductor (CMOS) sensor to detect a signal response
associated with the fluorescently labeled circulating tumor cells.
The optical sensor is connected to a controller configured to
control release of a chemotherapeutic agent from the second
reservoir positioned downstream from the optical sensor. The
implantable device also includes a transmitter for transmitting
data to an external receiver regarding the sensed levels of
circulating tumor cells.
[0240] The first reservoir of the device contains a
fluorescein-containing fluorescent label conjugated to an
oligonucleotide aptamer binding component. The aptamer selectively
binds to the epithelial cell-cell adhesion molecule (EpCAM)
expressed on circulating metastatic tumor cells of epithelial cell
origin. Fluorescein has a maximal absorbance at a wavelength of 494
nm and maximal emission at a wavelength of 521 nm (from, e.g.,
Molecular Probe--Invitrogen, Carlsbad, Calif.). Fluorescein is
conjugated to a binding component, e.g., an aptamer. Aptamers
directed against EpCAM are isolated from a large library of
10.sup.14 to 10.sup.15 random oligonucleotide sequences using an
iterative in vitro selection procedure termed "systematic evolution
of ligands by exponential enrichment" (SELEX). See, e.g., Cao, et
al., Current Proteomics 2:31-40, 2005; Proske, et al., Appl.
Microbiol. Biotechnol. 69:367-374, 2005; Jayasena Clin. Chem.
45:1628-1650, 1999, which are incorporated herein by reference. The
3-prime end of the EpCAM-specific aptamer is modified with an amino
group during chemical synthesis using a 3'-amino-modifier C7-CPG
(from, Glen Research, Sterling, Va.). Fluorescein is conjugated to
the amine-modified aptamer using a fluorescein-reactive dye
containing a succinimidyl ester moiety that reacts efficiently with
primary amines (e.g., 5-carboxyfluorescein, succinimidyl ester,
from Molecular Probes--Invitrogen, Carlsbad, Calif.).
[0241] The fluorescein-labeled aptamer is further modified with a
quenching molecule to create a FRET (fluorescence resonance energy
transfer) pair that will only fluoresce in response to binding a
target circulating tumor cell. An appropriate quencher to pair with
fluorescein is DABCYL acid
(4-(dimethylaminoazo)benzene-4-carboxylic acid). DABCYL is added to
the 5-prime end of the fluorescein-labeled aptamer during chemical
synthesis using 5'-DABCYL phosphoramidite
(6-(N-4'-carboxy-4-(dimethylamino)
azobenzene)-aminohexyl-1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidi-
te; from, e.g., Glen Research, Sterling, Va.). The aptamer is
configured such that in the absence of binding a circulating
metastatic tumor cell, fluorescein and DABCYL attached to the
aptamer are in close proximity and little to no fluorescence is
detected upon excitation. In response to binding a circulating
tumor cell, a conformational change occurs in the aptamer,
separating fluorescein and DABCYL and allowing fluorescence
associated with fluorescein to be detected.
[0242] The fluorescein/DABCYL labeled EpCAM aptamer is incorporated
into the first reservoir of the device and is released at constant
rate over time into the peripheral blood. In this example, the
EpCAM aptamer is formulated with poly (DL-lactide-co-glycolide)
(PLGA) into nanoparticles and coated onto some fraction of the
stent surface. See, e.g., Nakano, et al., J. Am. Coll. Cardiol.
Intv., 2:277-283, 2009, which is incorporated herein by reference.
Briefly, the fluorescein/DABCYL labeled EpCAM aptamer is
encapsulated into cationic PLGA nanopartices using an emulsion
solvent diffusion method. Electrodeposition is used to coat the
stent with the PLGA nanoparticles containing the fluorescein/DABCYL
labeled EpCAM aptamer. See, e.g., Htay &, Lui, Vasc. Health
Risk Management, 1:263-276, 2005, which is incorporated herein by
reference. While release of fluorescein/DABCYL labeled EpCAM into
the circulation occurs upstream of the optical sensor, it is
anticipated that interaction of the labeled aptamer with a
circulating tumor cell will occur elsewhere in the circulation, but
this interaction is ultimately detected when the labeled
circulating tumor cell passes through the implantable device.
[0243] Fluorescence associated with the fluorescein/DABCYL EpCAM
aptamer bound to a circulating metastatic tumor cell is detected
using an optical sensor. The optical sensor consists of a one-chip
sensing device designed for real time in vivo imaging and including
a complementary metal-oxide semiconductor (CMOS) sensor, a
blue-light emitting diode, and a color filter. See, e.g., Tamura,
et al. J. Neurosci. Methods 173:114-120, 2008; Ng, et al., J.
Neurosci. Methods 156:23-30, 2006, which are incorporated herein by
reference. The one-chip sensing device including the CMOS sensor is
incorporated into the stent. See, e.g., U.S. Patent Application
2009/0298704, which is incorporated herein by reference. In
response to sensing fluorescence associated with a circulating
tumor cell a signal is sent to the second reservoir to initiate
release of a chemotherapeutic agent. In addition, a signal
indicating detection of each circulating tumor cell is sent
wirelessly to an external receiver. See, e.g., U.S. Patent
Application 2009/0043183, which is incorporated herein by
reference. The external receiver is incorporated into a patch worn
by the subject. The data recorded in the patch is periodically
analyzed by the subject's physician to monitor the number of
circulating tumor cells detected over a given time frame, including
days and weeks.
[0244] The optical sensor is wired to the second reservoir
containing a chemotherapeutic agent, e.g., doxorubicin. In response
to detecting a fluorescent circulating tumor cell, the optical
sensor sends a signal to the second reservoir to trigger release of
the doxorubicin. The second reservoir is a microfabricated array of
micro-reservoirs incorporated into the stent. Each micro-reservoir
contains a measured bolus of doxorubicin. Each micro-reservoir is
sealed by a voltage sensitive metal foil responsive to an
electrical signal. See, e.g., Grayson, et al., Proc. IEEE, 92:6-21,
2004; which is incorporated herein by reference. Electrical signals
from the optical sensor trigger disintegration of the metal foil
and release of the enclosed bolus of doxorubicin.
[0245] The subject's physician may choose to add additional
medications to the subject's treatment regimen depending upon the
number of circulating metastatic tumor cells detected in the
subject's circulation. Additional medications can include
additional chemotherapy and/or radiation therapy. The implantable
device can continue to be used before and after these additional
treatment options to monitor for the presence of circulating
metastatic tumor cells and assess efficacy of the treatment
regimen.
Example 2
Device Including Sensor for Detecting One or More Target
Circulating Tumor Cells and for Binding and Altering Circulating
Tumor Cells for Treatment of a Neoplastic Disease or Condition
[0246] Referring to FIG. 4, depicted is a partial diagrammatic
cross-sectional view of an illustrative embodiment of an
implantable device 400 for treating a neoplastic disease including
a stent 410 including a lumen 415 configured for fluid flow; a
first reservoir 420 containing a source of fluorescent label
configured to bind circulating tumor cells in the peripheral blood
of a subject; an optical sensor 430 including a light emitting
diode 440 configured to provide energy to elicit a signal response
associated with fluorescently labeled circulating tumor cells and a
CMOS sensor 450 configured to detect signal responses associated
with fluorescently-labeled circulating tumor cells; and a second
reservoir 460 in communication with the optical sensor 430 through
a wired connection 470 and containing a chemotherapeutic agent
configured to cause apoptosis or necrosis of the circulating
metastatic tumor cell.
Example 3
Device Including Source for Labeling and Imaging Target
Inflammatory Mediators and for Ablating Inflammatory Mediators for
Treatment of an Inflammatory Condition or Disease
[0247] An implantable device is described for controlling elevated
levels of eosinophils in a subject experiencing an inflammatory
condition, e.g., asthma, by selectively sensing and controllably
binding and inducing apoptosis of the eosinophils. The components
of the implantable device are incorporated into an artificial
multi-lumen bypass graft surgically placed in a large artery, e.g.,
the coronary artery, of the subject. The components of the
implantable device include a first reservoir containing a
fluorescent label to bind and emit a fluorescent signal in response
to binding eosinophils in the blood of a subject; an optical sensor
including a set of light emitting diodes (LEDs) emitting light at a
specific wavelength to elicit a signal response associated with the
fluorescently labeled eosinophils; a metal-oxide-semiconductor
(CMOS) sensor to detect a signal response associated with elevated
levels of fluorescently labeled eosinophils; a controller
operationally linked to the optical sensor and configured to
control flow of the peripheral blood through the artificial
multi-lumen bypass graft in response to detecting elevated levels
of fluorescently-labeled eosinophils in the blood; and a second
reservoir including an apoptotic agent to selectively bind to and
inactivate the fluorescently-labeled eosinophils using an apoptotic
agent. The implantable device also includes a transmitter for
transmitting data to an external receiver regarding the sensed
levels of eosinophils in the blood of the subject.
[0248] The implantable device is incorporated into an artificial
multi-lumen bypass graft. The artificial multi-lumen bypass graft
is constructed of a biocompatible synthetic material, e.g.,
polyurethane, as described by Rashid, et al., FASEB J.,
22:2084-2089, 2008, which is incorporated herein by reference. The
artificial multi-lumen bypass graft includes one main flow path and
one side flow path branching from the main flow path. The first
reservoir containing the fluorescent label and the optical sensor
are incorporated into the main flow path and upstream of the
diversion point with the side flow path. Access to the side flow
path is controlled by a valve. The valve opens and closes in
response to signals sent from the optical sensor regarding the
sensed level of eosinophils. The side flow path is a multi-lumen
catheter composed of fluoropolymer (from Zeus, Orangeburg, S.C.)
through which the subject's blood is diverted. The multi-lumen
portion of the artificial graft constitutes the second reservoir of
the device and is configured to provide the blood components
including the fluorescently labeled eosinophils ample surface area
and exposure to an apoptotic agent.
[0249] The first reservoir of the device contains a Dapoxyl.RTM.
fluorescent label conjugated to an antibody binding component. The
antibody selectively binds to the IL-5 receptor expressed on the
surface of eosinophils. Dapoxyl.RTM. (benzenesulfonic acid,
4-[5-[4-(dimethylamino)phenyl]-2-oxazolyl) has an excitation maxima
at a wavelength of 370 nm and an emission maxima at a wavelength of
580 nm and is a solvent-sensitive fluorophore that does not
fluoresce well when exposed to an aqueous environment, e.g., blood
(from, e.g., Molecular Probe--Invitrogen, Carlsbad, Calif.).
Antibodies directed against the IL-5 receptor are available from
commercial sources (from, e.g., Sigma-Aldrich, St. Louis, Mo.). A
target responsive Dapoxyl.RTM.-antibody conjugate is constructed by
combining the IL-5 receptor antibody with the IL-5 receptor and the
complex is non-selectively labeled with Dapoxyl.RTM. near the
active site of the antibody. The IL-5 receptor antigen is removed,
leaving Dapoxyl.RTM. labeled antibody with poor fluorescence in
aqueous environments. Interaction of the IL-5 receptor antigen with
the Dapoxyl.RTM.-labeled antibody protects the label near the
active site from the aqueous environment and results in a large
blue shift in the emission wavelength as well as an increase in
fluorescence intensity. See, e.g., Brennan, J. Fluor. 9: 298-312,
1999, which is incorporated herein by reference.
[0250] The Dapoxyl.RTM.-labeled IL-5 receptor antibody is
incorporated into the first reservoir of the device and is released
at constant rate over time into the peripheral blood. The first
reservoir includes an infusion pump attached to the exterior of the
artificial multi-lumen bypass graft with flow access to the
internal lumen of the main flow path. The infusion pump injects the
labeled IL-5 receptor antibody into the blood flowing through the
main flow path of the device. The SynchroMed II.RTM. implantable
infusion pump (from Medtronic, Minneapolis, Minn.) is an example of
an implantable infusion pump.
[0251] Fluorescence associated with the Dapoxyl.RTM.-labeled IL-5
receptor antibody bound to eosinophils is detected using an optical
sensor. The optical sensor is positioned in the main flow path of
the artificial multi-lumen bypass graft and upstream of the
diversion point with the side flow path. The optical sensor
consists of a sensing device designed for real time in vivo imaging
and including a complementary metal-oxide semiconductor (CMOS)
sensor, an ultraviolet emitting diode, and a color filter
appropriate for capturing fluorescence associated with Dapoxyl.RTM.
excitation. See, e.g., Tamura, et al. J. Neurosci. Methods 173:
114-120, 2008; Ng, et al., J. Neurosci. Methods 156: 23-30, 2006,
which are incorporated herein by reference. The CMOS sensor is
configured to count the number of fluorescence "events" over a
period of time. Each fluorescence event represents an eosinophil
passing by the CMOS sensor. When the rate of fluorescence events
exceeds a threshold value indicative of excessive eosinophils in
circulation, the sensor sends a signal to the valve controlling
access to the side flow path. A signal is also sent wirelessly to
an external receiver indicating the presence of excessive
eosinophils in the subject's blood. See, e.g., U.S. Patent
Application 2009/0043183, which is incorporated herein by
reference. The data transmitted to the external receiver is
periodically analyzed by the subject's physician to monitor the
number of eosinophils detected over a given time frame, including
days and weeks.
[0252] In response to reaching a threshold of fluorescence events,
the controller associate with the CMOS sensor sends a signal to
open the valve and at least a portion of the peripheral blood
flowing through the artificial multi-lumen bypass graft is diverted
into the multi-lumens of the side flow path. The multi-lumens of
the side path are coated with 6'-sulfated sialyl-Lewis X, a
carbohydrate structure that binds to Siglec-8, an
immunoglobulin-like protein selectively expressed on the surface of
eosinophils. The interaction of Siglec-8 with 6'-sulfated
sialyl-Lewis X initiates caspase- and mitochondria-mediated
apoptosis of the eosinophils. See, e.g., Nutku-Bilir, et al., Am.
J. Respir. Cell. Mol. Biol., 38: 121-124, which is incorporated
herein by reference. Other cells in the blood lacking Siglec-8 are
not affected by the presence of 6'-sulfated sialyl-Lewis X and pass
through the side flow path and downstream to the main flow
path.
[0253] The controller is configured to adjust access to the side
flow path to achieve a target value of the fluorescently labeled
eosinophils in the peripheral blood of the subject. The controller
calculates the level of eosinophils in the peripheral blood based
on input from the optical sensor, e.g., fluorescence events, and
compares these data with target values, e.g., desired
concentrations of eosinophils. The number of eosinophils in a
normal human subject ranges from about 40 cells to about 400 cells
per microliter of blood. More than 500 cells per microliter of
blood is considered eosinophilia in adults. In some instances, the
target value for eosinophils is that observed in a normal subject
not experiencing an inflammatory disease. In other instances, the
target value for eosinophils may represent a reduction of at least
20%, at least 40%, at least 60%, at least 80%, or at least 100%
relative to the current level of eosinophils in the peripheral
blood of the subject.
Example 4
Device Including Source for Labeling and Imaging Target
Inflammatory Mediators and for Ablating Inflammatory Mediators for
Treatment of an Inflammatory Condition or Disease
[0254] Referring to FIG. 5A, depicted is a partial diagrammatic
view of an illustrative embodiment of an implantable device 500 for
modulating the levels of eosinophils in a subject including an
artificial multi-lumen bypass graft 510 configured for fluid flow.
The artificial multi-lumen bypass graft 510 is constructed of a
biocompatible synthetic material, e.g., polyurethane, as described
by Rashid, et al., FASEB J., 22: 2084-2089, 2008, which is
incorporated herein by reference. The artificial multi-lumen bypass
graft 510 includes a lumen including a main flow path 520 and a
side flow path 530 branching from the main flow path 520. The side
flow path 530 is separated from the main flow path 520 by a valve
540. The artificial multi-lumen bypass graft 510 further includes a
first reservoir 550 with an access port 560 to the main flow path
520 and containing a fluorescent label; a light emitting diode 570
configured to provide energy to elicit a signal response associated
with fluorescently-labeled eosinophils; and a CMOS sensor 580
configured to detect signal responses associated with
fluorescently-labeled eosinophils. Valve 540 controls access to
side flow path 530 and opens or closes in response to signals sent
from CMOS sensor 580 regarding the sensed level of
fluorescently-labeled eosinophils. The side flow path 530 is a
multi-lumen catheter composed of fluoropolymer (from Zeus,
Orangeburg, S.C.) through which the subject's blood is diverted.
Referring to FIG. 5B, depicted is a cross-section A through side
flow path 530 of FIG. 5A. Side flow path 530 contains multiple
parallel lumens 590. Each of the multiple parallel lumens 590
constitutes a second reservoir of the implantable device 500. The
multiple parallel lumens 590 are configured to provide the blood
components including the fluorescently labeled eosinophils ample
surface area and exposure to an apoptotic agent, e.g., 6'-sulfated
sialyl-Lewis X, to destroy the fluorescently labeled
eosinophils.
[0255] Each recited range includes all combinations and
sub-combinations of ranges, as well as specific numerals contained
therein.
[0256] All publications and patent applications cited in this
specification are herein incorporated by reference to the extent
not inconsistent with the description herein and for all purposes
as if each individual publication or patent application were
specifically and individually indicated to be incorporated by
reference for all purposes.
[0257] The state of the art has progressed to the point where there
is little distinction left between hardware and software
implementations of aspects of systems; the use of hardware or
software is generally (but not always, in that in certain contexts
the choice between hardware and software can become significant) a
design choice representing cost vs. efficiency tradeoffs. There are
various vehicles by which processes and/or systems and/or other
technologies described herein can be effected (e.g., hardware,
software, and/or firmware), and that the preferred vehicle will
vary with the context in which the processes and/or systems and/or
other technologies are deployed. For example, if an implementer
determines that speed and accuracy are paramount, the implementer
may opt for a mainly hardware and/or firmware vehicle;
alternatively, if flexibility is paramount, the implementer may opt
for a mainly software implementation; or, yet again alternatively,
the implementer may opt for some combination of hardware, software,
and/or firmware. Hence, there are several possible vehicles by
which the processes and/or devices and/or other technologies
described herein may be effected, none of which is inherently
superior to the other in that any vehicle to be utilized is a
choice dependent upon the context in which the vehicle will be
deployed and the specific concerns (e.g., speed, flexibility, or
predictability) of the implementer, any of which may vary. Optical
aspects of implementations will typically employ optically-oriented
hardware, software, and/or firmware.
[0258] In a general sense the various aspects described herein
which can be implemented, individually and/or collectively, by a
wide range of hardware, software, firmware, or any combination
thereof can be viewed as being composed of various types of
"electrical circuitry." "Electrical circuitry" includes, but is not
limited to, electrical circuitry having at least one discrete
electrical circuit, electrical circuitry having at least one
integrated circuit, electrical circuitry having at least one
application specific integrated circuit, electrical circuitry
forming a general purpose computing device configured by a computer
program (e.g., a general purpose computer configured by a computer
program which at least partially carries out processes and/or
devices described herein, or a microprocessor configured by a
computer program which at least partially carries out processes
and/or devices described herein), electrical circuitry forming a
memory device (e.g., forms of random access memory), and/or
electrical circuitry forming a communications device (e.g., a
modem, communications switch, or optical-electrical equipment). The
subject matter described herein may be implemented in an analog or
digital fashion or some combination thereof.
[0259] The herein described components (e.g., steps), devices, and
objects and the description accompanying them are used as examples
for the sake of conceptual clarity and that various configuration
modifications using the disclosure provided herein are within the
skill of those in the art. Consequently, the specific examples set
forth and the accompanying description are intended to be
representative of their more general classes. In general, use of
any specific example herein is also intended to be representative
of its class, and the non-inclusion of such specific components
(e.g., steps), devices, and objects herein should not be taken as
indicating that limitation is desired.
[0260] With respect to the use of substantially any plural or
singular terms herein, the reader can translate from the plural to
the singular or from the singular to the plural as is appropriate
to the context or application. The various singular/plural
permutations are not expressly set forth herein for sake of
clarity.
[0261] The herein described subject matter sometimes illustrates
different components contained within, or connected with, different
other components. It is to be understood that such depicted
architectures are merely examples, and that in fact many other
architectures can be implemented which achieve the same
functionality. In a conceptual sense, any arrangement of components
to achieve the same functionality is effectively "associated" such
that the desired functionality is achieved. Hence, any two
components herein combined to achieve a particular functionality
can be seen as "associated with" each other such that the desired
functionality is achieved, irrespective of architectures or
intermedial components. Likewise, any two components so associated
can also be viewed as being "operably connected," or "operably
coupled," to each other to achieve the desired functionality, and
any two components capable of being so associated can also be
viewed as being "operably couplable," to each other to achieve the
desired functionality. Specific examples of operably couplable
include but are not limited to physically mateable or physically
interacting components or wirelessly interactable or wirelessly
interacting components or logically interacting or logically
interactable components.
[0262] While particular aspects of the present subject matter
described herein have been shown and described, changes and
modifications may be made without departing from the subject matter
described herein and its broader aspects and, therefore, the
appended claims are to encompass within their scope all such
changes and modifications as are within the true spirit and scope
of the subject matter described herein. Furthermore, it is to be
understood that the invention is defined by the appended claims. It
will be understood that, in general, terms used herein, and
especially in the appended claims (e.g., bodies of the appended
claims) are generally intended as "open" terms (e.g., the term
"including" should be interpreted as "including but not limited
to," the term "having" should be interpreted as "having at least,"
the term "includes" should be interpreted as "includes but is not
limited to," etc.). It will be further understood that if a
specific number of an introduced claim recitation is intended, such
an intent will be explicitly recited in the claim, and in the
absence of such recitation no such intent is present. For example,
as an aid to understanding, the following appended claims may
contain usage of the introductory phrases "at least one" and "one
or more" to introduce claim recitations. However, the use of such
phrases should not be construed to imply that the introduction of a
claim recitation by the indefinite articles "a" or "an" limits any
particular claim containing such introduced claim recitation to
inventions containing only one such recitation, even when the same
claim includes the introductory phrases "one or more" or "at least
one" and indefinite articles such as "a" or "an"; the same holds
true for the use of definite articles used to introduce claim
recitations. In addition, even if a specific number of an
introduced claim recitation is explicitly recited, such recitation
should typically be interpreted to mean at least the recited number
(e.g., the bare recitation of "two recitations," without other
modifiers, typically means at least two recitations, or two or more
recitations). Furthermore, in those instances where a convention
analogous to "at least one of A, B, and C, etc." is used, in
general such a construction is intended in the sense one having
skill in the art would understand the convention (e.g., "a system
having at least one of A, B, and C" would include but not be
limited to systems that have A alone, B alone, C alone, A and B
together, A and C together, B and C together, or A, B, and C
together, etc.). In those instances where a convention analogous to
"at least one of A, B, or C, etc." is used, in general such a
construction is intended in the sense one having skill in the art
would understand the convention (e.g., "a system having at least
one of A, B, or C" would include but not be limited to systems that
have A alone, B alone, C alone, A and B together, A and C together,
B and C together, or A, B, and C together, etc.). Virtually any
disjunctive word and/or phrase presenting two or more alternative
terms, whether in the description, claims, or drawings, should be
understood to contemplate the possibilities of including one of the
terms, either of the terms, or both terms. For example, the phrase
"A or B" will be understood to include the possibilities of "A" or
"B" or "A and B."
[0263] While various aspects and embodiments have been disclosed
herein, other aspects and embodiments will be apparent to those
skilled in the art. The various aspects and embodiments disclosed
herein are for purposes of illustration and are not intended to be
limiting, with the true scope and spirit being indicated by the
following claims.
* * * * *
References