U.S. patent application number 15/720329 was filed with the patent office on 2018-01-25 for sensor head for use with implantable devices.
The applicant listed for this patent is DexCom, Inc.. Invention is credited to James H. Brauker, Rathbun K. Rhodes, Mark C. Shults, Mark A. Tapsak.
Application Number | 20180024086 15/720329 |
Document ID | / |
Family ID | 25437709 |
Filed Date | 2018-01-25 |
United States Patent
Application |
20180024086 |
Kind Code |
A1 |
Rhodes; Rathbun K. ; et
al. |
January 25, 2018 |
SENSOR HEAD FOR USE WITH IMPLANTABLE DEVICES
Abstract
The present invention provides a sensor head for use in an
implantable device that measures the concentration of an analyte in
a biological fluid which includes: a non-conductive body; a working
electrode, a reference electrode and a counter electrode, wherein
the electrodes pass through the non-conductive body forming an
electrochemically reactive surface at one location on the body and
forming an electronic connection at another location on the body,
further wherein the electrochemically reactive surface of the
counter electrode is greater than the surface area of the working
electrode; and a multi-region membrane affixed to the nonconductive
body and covering the working electrode, reference electrode and
counter electrode. In addition, the present invention provides an
implantable device including at least one of the sensor heads of
the invention and methods of monitoring glucose levels in a host
utilizing the implantable device of the invention.
Inventors: |
Rhodes; Rathbun K.;
(Madison, WI) ; Tapsak; Mark A.; (Bloomsburg,
PA) ; Brauker; James H.; (Addison, MI) ;
Shults; Mark C.; (Madison, WI) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
DexCom, Inc. |
San Diego |
CA |
US |
|
|
Family ID: |
25437709 |
Appl. No.: |
15/720329 |
Filed: |
September 29, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15059086 |
Mar 2, 2016 |
9804114 |
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15720329 |
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13943622 |
Jul 16, 2013 |
9328371 |
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15059086 |
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12260017 |
Oct 28, 2008 |
8509871 |
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13943622 |
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11021162 |
Dec 22, 2004 |
7471972 |
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12260017 |
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09916711 |
Jul 27, 2001 |
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11021162 |
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Current U.S.
Class: |
204/403.14 ;
600/345 |
Current CPC
Class: |
C12Q 1/006 20130101;
A61B 5/14532 20130101; A61B 5/14865 20130101; C12Q 1/002 20130101;
G01N 33/48785 20130101; G01N 27/3272 20130101; G01N 33/48707
20130101 |
International
Class: |
G01N 27/327 20060101
G01N027/327; C12Q 1/00 20060101 C12Q001/00; A61B 5/1486 20060101
A61B005/1486; G01N 33/487 20060101 G01N033/487; A61B 5/145 20060101
A61B005/145 |
Claims
1. A sensor for use in a glucose measuring device, the sensor
comprising: a first electrode, a second electrode, and a
non-conductive body located between the first electrode and the
second electrode, wherein the first electrode and the second
electrode each form an electrochemically reactive surface at one
end of the sensor and an electronic connection at another end of
the sensor; and a multi-region membrane covering the first
electrode and the second electrode, wherein the multi-region
membrane comprises an immobilized enzyme domain comprising an
enzyme in at least a portion thereof, wherein the multi-region
membrane comprises a glucose exclusion domain that is permeable to
oxygen and interferes with glucose transport across said membrane,
and wherein said glucose exclusion domain does not cover the
working electrode.
2. The sensor of claim 1, wherein the multi-region membrane further
comprises an interference domain more proximal to said
electrochemically reactive surfaces than said glucose exclusion
domain.
3. The sensor of claim 1, wherein the multi-region membrane further
comprises a resistance domain
4. The sensor of claim 1, wherein the multi-region membrane further
comprises an immobilized enzyme domain more proximal to said
electrochemically reactive surfaces than said glucose exclusion
domain.
5. The sensor of claim 1, wherein the multi-region membrane
comprises a hydrogel domain adjacent to said electrochemically
reactive surfaces.
6. A sensor for use in a glucose measuring device, the sensor
comprising: a first electrode, a second electrode, and a
non-conductive body located between the first electrode and the
second electrode, wherein the first electrode and the second
electrode each form an electrochemically reactive surface at one
end of the sensor and an electronic connection at another end of
the sensor; and a multi-region membrane covering the first
electrode and the second electrode, wherein the multi-region
membrane comprises an immobilized enzyme domain comprising an
enzyme in at least a portion thereof, wherein the multi-region
membrane comprises a portion positioned over only said counter
electrode that reduces the consumption of oxygen above said counter
electrode.
7. The sensor of claim 6, wherein the portion comprises
silicone.
8. The sensor head of claim 6, wherein an active enzyme is
positioned only over the working electrode.
9. The sensor of claim 6, wherein an inactive enzyme is positioned
only over at least one of the first electrode and the second
electrode.
Description
INCORPORATION BY REFERENCE TO RELATED APPLICATIONS
[0001] Any and all priority claims identified in the Application
Data Sheet, or any correction thereto, are hereby incorporated by
reference under 37 CFR 1.57. This application is a continuation of
U.S. Application No. 15/059,086, filed March 2, 2016, which is a
continuation of U.S. application Ser. No. 13/943,622, filed Jul.
16, 2013, now U.S. Pat. No. 9,328,371, which is a continuation of
U.S. application Ser. No. 12/260,017, filed on Oct. 28, 2008, now
U.S. Pat. No. 8,509,871, which is a division of U.S. application
Ser. No. 11/021,162, filed Dec. 22, 2004, now U.S. Pat. No.
7,471,972, which is a continuation of U.S. application Ser. No.
09/916,711, filed Jul. 27, 2001, now abandoned. Each of the
aforementioned applications is incorporated by reference herein in
its entirety, and each is hereby expressly made a part of this
specification.
FIELD OF THE INVENTION
[0002] The present invention relates generally to novel sensor
heads utilized with implantable devices, devices including these
sensor heads and methods for determining analyte levels using these
implantable devices. More particularly, the invention relates to
sensor heads, implantable devices including these sensor heads and
methods for monitoring glucose levels in a biological fluid using
these devices.
BACKGROUND OF THE INVENTION
[0003] Amperometric electrochemical sensors require a counter
electrode to balance the current generated by the species being
measured at the working electrode. In the case of a glucose oxidase
based glucose sensor, the species being measured at the working
electrode is H.sub.2O.sub.2. Glucose oxidase catalyzes the
conversion of oxygen and glucose to hydrogen peroxide and gluconate
according to the following reaction:
Glucose+O.sub.2.fwdarw.Gluconate+H.sub.2O.sub.2
[0004] Because for each glucose molecule metabolized, there is a
proportional change in the product H.sub.2O.sub.2, one can monitor
the change in H.sub.2O.sub.2 to determine glucose concentration.
Oxidation of H.sub.2O.sub.2 by the working electrode is balanced by
reduction of ambient oxygen, enzyme generated H.sub.2O.sub.2, or
other reducible species at the counter electrode. In vivo glucose
concentration may vary from about one hundred times or more that of
the oxygen concentration. Consequently, oxygen becomes a limiting
reactant in the electrochemical reaction and when insufficient
oxygen is provided to the sensor, the sensor will be unable to
accurately measure glucose concentration. Those skilled in the art
have come to interpret oxygen limitations resulting in depressed
function as being a problem of availability of oxygen to the
enzyme.
[0005] As shown in FIG. 1, the sensor head 10 includes a working
electrode 21 (anode), counter electrode 22 (cathode), and reference
electrode 20 which are affixed to the head by both brazing 26 the
electrode metal to the ceramic and potting with epoxy 28. The
working electrode 21 (anode) and counter-electrode 22 (cathode) of
a glucose oxidase-based glucose sensor head 10 require oxygen in
different capacities. Prior art teaches an enzyme-containing
membrane that resides above an amperometric electrochemical sensor.
In FIG. 1, region 32 includes an immobilized enzyme, i.e. glucose
oxidase. Within the enzyme layer above the working electrode 21,
oxygen is required for the production of H.sub.2O.sub.2 from
glucose. The H.sub.2O.sub.2 produced from the glucose oxidase
reaction further reacts at surface 21a of working electrode 21 and
produces two electrons. The products of this reaction are two
protons (2H.sub.+), two electrons (2e.sup.-), and one oxygen
molecule (O.sub.2) (Fraser, D. M. "An Introduction to In Vivo
Biosensing: Progress and problems." In "Biosensors and the Body,"
D. M. Fraser, ed., 1997, pp. 1-56 John Wiley and Sons, New York).
In theory, the oxygen concentration near the working electrode 21,
which is consumed during the glucose oxidase reaction, is
replenished by the second reaction at the working electrode.
Therefore, the net consumption of oxygen is zero. In practice,
neither all of the H.sub.2O.sub.2 produced by the enzyme diffuses
to the working electrode surface nor does all of the oxygen
produced at the electrode diffuse to the enzyme domain.
[0006] With further reference to FIG. 1, the counter electrode 22
utilizes oxygen as an electron acceptor. The most likely reducible
species for this system are oxygen or enzyme generated peroxide
(Fraser, D. M. supra). There are two main pathways by which oxygen
may be consumed at the counter electrode 22. These are a
four-electron pathway to produce hydroxide and a two-electron
pathway to produce hydrogen peroxide. The two-electron pathway is
shown in FIG. 1. Oxygen is further consumed above the counter
electrode by the glucose oxidase in region 32. Due to the oxygen
consumption by both the enzyme and the counter electrode, there is
a net consumption of oxygen at the surface 22a of the counter
electrode. Theoretically, in the domain of the working electrode
there is significantly less net loss of oxygen than in the region
of the counter electrode. In addition, there is a close correlation
between the ability of the counter electrode to maintain current
balance and sensor function. Taken together, it appears that
counter electrode function becomes limited before the enzyme
reaction becomes limited when oxygen concentration is lowered.
[0007] Those practicing in the field of implantable glucose oxidase
sensors have focused on improving sensor function by increasing the
local concentration of oxygen in the region of the working
electrode. (Fraser, D. M. supra).
[0008] We have observed that in some cases, loss of glucose oxidase
sensor function may not be due to a limitation of oxygen in the
enzyme layer near the working electrode, but may instead be due to
a limitation of oxygen at the counter electrode. In the presence of
increasing glucose concentrations, a higher peroxide concentration
results, thereby increasing the current at the working electrode.
When this occurs, the counter electrode limitation begins to
manifest itself as this electrode moves to increasingly negative
voltages in the search for reducible species. When a sufficient
supply of reducible species, such as oxygen, are not available, the
counter electrode voltage reaches a circuitry limit of -0.6V
resulting in compromised sensor function (see FIG. 3).
[0009] FIG. 3 shows simultaneous measurement of counter-electrode
voltage and sensor output to glucose levels from a glucose sensor
implanted subcutaneously in a canine host. It can be observed that
as glucose levels increase, the counter electrode voltage
decreases. When the counter electrode voltage reaches -0.6V, the
signal to noise ratio increases significantly. This reduces the
accuracy of the device. FIG. 4 shows a further example of another
glucose sensor in which the counter-electrode reaches the circuitry
limit. Again, once the counter electrode reaches -0.6V, the
sensitivity and/or signal to noise ratio of the device is
compromised. In both of these examples, glucose levels reached
nearly 300 mg/dl. However, in FIG. 3 the sensor showed a greater
than three-fold higher current output than the sensor in FIG. 4.
These data suggest that there may be a limitation of reducible
species at the counter electrode, which may limit the sensitivity
of the device as the glucose levels increase. In contrast, FIG. 5
shows a glucose sensor in which the counter electrode voltage did
not reach -0.6V. In FIG. 5 it can be observed that the sensor was
able to maintain a current balance between the working and counter
electrodes, thereby enabling accurate measurements throughout the
course of the experiment. The results shown in FIGS. 3, 4 and 5 led
the present inventors to postulate that by keeping the counter
electrode from reaching the circuitry limit, one could maintain
sensitivity and accuracy of the device.
[0010] Two approaches have been utilized by others to relieve the
counter electrode limitation described above. The first approach
involves the widening of the potential range over which the counter
electrode can move in the negative direction to avoid reaching
circuitry limitations. Unfortunately, this approach increases
undesirable products that are produced at lower potentials. One
such product, hydrogen, may form at the counter electrode, which
may then diffuse back to the working electrode. This may contribute
to additional current resulting in erroneously high glucose
concentration readings. Additionally, at these increasingly
negative potentials, the probability of passivating or poisoning
the counter electrode greatly increases. This effectively reduces
the counter electrode surface area requiring a higher current
density at the remaining area to maintain current balance.
Furthermore, increased current load increases the negative
potentials eventually resulting in electrode failure.
[0011] The second approach is utilizing the metal case of the
device as a counter electrode (see U.S. Pat. No. 4,671,288, Gough
or U.S. Pat. No. 5,914,026, Blubaugh). This provides an initial
excess in surface area which is expected to serve the current
balancing needs of the device over its lifetime. However, when the
counter electrode reaction is a reduction reaction, as in Blubaugh,
the normally present metal oxide layer will be reduced to bare
metal over time leaving the surface subject to corrosion,
poisoning, and eventual cascade failure. This problem is magnified
when considering the various constituents of the body fluid that
the metal casing is exposed to during in vivo use. To date, there
has been no demonstration of long-term performance of such a device
with this counter electrode geometry.
[0012] Consequently, there is a need for a sensor that will provide
accurate analyte measurements, that reduces the potential for
cascade failure due to increasing negative potentials, corrosion
and poisoning, and that will function effectively and efficiently
in low oxygen concentration environments.
SUMMARY OF THE INVENTION
[0013] In one aspect of the present invention, a sensor head for
use in a device that measures the concentration of an analyte in a
biological fluid is provided that includes a non-conductive body; a
working electrode, a reference electrode and a counter electrode,
wherein the electrodes pass through the non-conductive body forming
an electrochemically reactive surface at one location on the body
and forming an electronic connection at another location on the
body, and further wherein the electrochemically reactive surface of
the counter electrode is greater than the surface area of the
working electrode; and a multi-region membrane affixed to the
nonconductive body and covering the working electrode, reference
electrode and counter electrode.
[0014] In another aspect of the present invention, a sensor head
for use in an implantable analyte measuring device is provided
which includes the same sensor head components as those described
above.
[0015] The sensor heads of the present invention include a
multi-region membrane that controls the number of species that are
able to reach the surface of the electrodes. In particular, such a
membrane allows the passage of desired substrate molecules (e.g.
oxygen and glucose) and rejects other larger molecules that may
interfere with accurate detection of an analyte. The sensor heads
of the present invention also provide a larger counter electrode
reactive surface that balances the current between the working and
counter electrodes, thereby minimizing negative potential extremes
that may interfere with accurate analyte detection.
[0016] In another aspect of the present invention, an implantable
device for measuring an analyte in a biological fluid is provided
including at least one of the sensor heads described above. In
still another aspect of the present invention, a method of
monitoring glucose levels is disclosed which includes the steps of
providing a host, and an implantable device as provided above and
implanting the device in the host.
[0017] Further encompassed by the invention is a method of
measuring glucose in a biological fluid including the steps of
providing a host and a implantable device described above, which
includes a sensor head capable of accurate continuous glucose
sensing; and implanting the device in the host.
[0018] The sensor head, membrane architectures, devices and methods
of the present invention allow for the collection of continuous
information regarding desired analyte levels (e.g. glucose). Such
continuous information enables the determination of trends in
glucose levels, which can be extremely important in the management
of diabetic patients.
DEFINITIONS
[0019] In order to facilitate an understanding of the present
invention, a number of terms are defined below.
[0020] The term "sensor head" refers to the region of a monitoring
device responsible for the detection of a particular analyte. The
sensor head generally comprises a non-conductive body, a working
electrode (anode), a reference electrode and a counter electrode
(cathode) passing through and secured within the body forming an
electrochemically reactive surface at one location on the body and
an electronic connective means at another location on the body, and
a multi-region membrane affixed to the body and covering the
electrochemically reactive surface. The counter electrode has a
greater electrochemically reactive surface area than the working
electrode. During general operation of the sensor a biological
sample (e.g., blood or interstitial fluid) or a portion thereof
contacts (directly or after passage through one or more membranes
or domains) an enzyme (e.g., glucose oxidase); the reaction of the
biological sample (or portion thereof) results in the formation of
reaction products that allow a determination of the analyte (e.g.
glucose) level in the biological sample. In preferred embodiments
of the present invention, the multi-region membrane further
comprises an enzyme domain, and an electrolyte phase (i.e., a
free-flowing liquid phase comprising an electrolyte-containing
fluid described further below).
[0021] The term "analyte" refers to a substance or chemical
constituent in a biological fluid (e.g., blood, interstitial fluid,
cerebral spinal fluid, lymph fluid or urine) that can be analyzed.
A preferred analyte for measurement by the sensor heads, devices
and methods of the present invention is glucose.
[0022] The term "electrochemically reactive surface" refers to the
surface of an electrode where an electrochemical reaction takes
place. In the case of the working electrode, the hydrogen peroxide
produced by the enzyme catalyzed reaction of the analyte being
detected reacts creating a measurable electronic current (e.g.
detection of glucose analyte utilizing glucose oxidase produces
H.sub.2O.sub.2 peroxide as a by product, H.sub.2O.sub.2 reacts with
the surface of the working electrode producing two protons
(2H.sub.+), two electrons (2e.sup.-) and one molecule of oxygen
(O.sub.2) which produces the electronic current being detected). In
the case of the counter electrode, a reducible species, e.g.
O.sub.2 is reduced at the electrode surface in order to balance the
current being generated by the working electrode.
[0023] The term "electronic connection" refers to any electronic
connection known to those in the art that may be utilized to
interface the sensor head electrodes with the electronic circuitry
of a device such as mechanical (e.g., pin and socket) or
soldered.
[0024] The term "domain" refers to regions of the membrane of the
present invention that may be layers, uniform or non-uniform
gradients (e.g. anisotropic) or provided as portions of the
membrane.
[0025] The term "multi-region membrane" refers to a permeable
membrane that may be comprised of two or more domains and
constructed of biomaterials of a few microns thickness or more
which are permeable to oxygen and may or may not be permeable to
glucose. One of the membranes may be placed over the sensor body to
keep host cells (e.g., macrophages) from gaining proximity to, and
thereby damaging, the enzyme membrane or forming a barrier cell
layer and interfering with the transport of analyte across the
tissue-device interface.
[0026] The phrase "distant from" refers to the spatial relationship
between various elements in comparison to a particular point of
reference. For example, some embodiments of a biological fluid
measuring device comprise a multi-region membrane that may be
comprised of a number of domains. If the electrodes of the sensor
head are deemed to be the point of reference, and one of the
multi-region membrane domains is positioned farther from the
electrodes, than that domain is distant from the electrodes.
[0027] The term "oxygen antenna domain" and the like refers to a
domain composed of a material that has higher oxygen solubility
than aqueous media so that it concentrates oxygen from the
biological fluid surrounding the biointerface membrane. The domain
can then act as an oxygen reservoir during times of minimal oxygen
need and has the capacity to provide on demand a higher oxygen
gradient to facilitate oxygen transport across the membrane. This
enhances function in the enzyme reaction domain and at the counter
electrode surface when glucose conversion to hydrogen peroxide in
the enzyme domain consumes oxygen from the surrounding domains.
Thus, this ability of the oxygen antenna domain to apply a higher
flux of oxygen to critical domains when needed improves overall
sensor function.
[0028] The term "solid portions" and the like refer to a material
having a structure that may or may not have an open-cell
configuration but in either case prohibits whole cells from
traveling through or residing within the material.
[0029] The term "substantial number" refers to the number of
cavities or solid portions having a particular size within a domain
in which greater than 50 percent of all cavities or solid portions
are of the specified size, preferably greater than 75 percent and
most preferably greater than 90 percent of the cavities or solid
portions have the specified size.
[0030] The term "co-continuous" and the like refers to a solid
portion wherein an unbroken curved line in three dimensions exists
between any two points of the solid portion.
[0031] The term "host" refers to both humans and animals.
[0032] The term "accurately" means, for example, 90% of measured
glucose values are within the "A" and "B" region of a standard
Clarke error grid when the sensor measurements are compared to a
standard reference measurement. It is understood that like any
analytical device, calibration, calibration validation and
recalibration are required for the most accurate operation of the
device.
[0033] The phrase "continuous glucose sensing" refers to the period
in which monitoring of plasma glucose concentration is continuously
performed, for example, about every 10 minutes.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] FIG. 1 Illustration of thermodynamically favored reactions
at the working electrode and counter electrode at the desired
voltage potentials.
[0035] FIG. 2A depicts a cross-sectional exploded view of a sensor
head of the present invention wherein the multi-region membrane
comprises three regions.
[0036] FIG. 2B depicts a cross-sectional exploded view of a sensor
head of the present invention wherein a portion of the second
membrane region does not cover the working electrode.
[0037] FIG. 2C depicts a cross-sectional exploded view of a sensor
head of the present invention which includes two distinct regions,
wherein the region adjacent the electrochemically reactive surfaces
includes a portion positioned over the counter electrode which
corresponds to a silicone domain.
[0038] FIG. 2D depicts a cross-sectional exploded view of a sensor
head of the present invention wherein an active enzyme of the
immobilized enzyme domain is positioned only over the working
electrode.
[0039] FIG. 2E depicts a cross-sectional exploded view of a sensor
head of the present invention wherein the enzyme positioned over
the counter electrode has been inactivated.
[0040] FIG. 2F depicts a cross-sectional exploded view of a sensor
head of the present invention wherein the membrane region
containing immobilized enzyme is positioned only over the working
electrode.
[0041] FIG. 3 Illustration of an implantable glucose sensor's
ability to measure glucose concentration during an infusion study
in a canine when the counter electrode voltage drops to the
electronic circuitry limit at approximately 0.75 hours wherein the
sensor current output reaches 2.50 nA.
[0042] FIG. 4 Illustration of an implantable glucose sensor's
ability to measure glucose concentration during an infusion study
in a canine when the counter electrode voltage drops to the
electronic circuitry limit after 0.5 hours wherein the sensor
current output reaches 0.50 nA.
[0043] FIG. 5 Illustration of an implantable glucose sensor's
ability to measure glucose concentration during an infusion study
in a canine when the counter electrode voltage is maintained above
the electronic circuitry limit.
[0044] FIG. 6A shows a schematic representation of a cylindrical
analyte measuring device including a sensor head according to the
present invention.
[0045] FIG. 6B is an exploded view of the sensor head of the device
shown in FIG. 6A.
[0046] FIG. 7 Graphical representation of the function of a device
of the present invention utilizing the multi-region membrane
architecture of FIG. 2B in vitro at 400 mg/dL glucose.
[0047] FIG. 8 depicts a cross-sectional exploded view of the
electrode and membrane regions of a prior sensor device where the
electrochemical reactive surface of the counter electrode is
substantially equal to the surface area of the working
electrode.
[0048] FIG. 9 Graphical representation of the counter electrode
voltage as a function of oxygen concentration at 400 mg/dL glucose
for sensor devices including the membrane shown in FIG. 8.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
[0049] In a preferred embodiment, the sensor heads, devices and
methods of the present invention may be used to determine the level
of glucose or other analytes in a host. The level of glucose is a
particularly important measurement for individuals having diabetes
in that effective treatment depends on the accuracy of this
measurement.
[0050] The present invention increases the effectiveness of counter
electrode function by a method that does not depend on increasing
the local concentration of oxygen. In a preferred embodiment, the
counter electrode has an electrochemical reactive surface area
greater than twice the surface area of the working electrode
thereby substantially increasing the electrodes ability to utilize
oxygen as a substrate. Further enhancement of the counter
electrode's activity may be achieved if the electrode were made of
gold. In a second preferred embodiment, the counter electrode has a
textured surface, with surface topography that increases the
surface area of the electrode while the diameter of the electrode
remains constant. In a third preferred embodiment, the proximity of
the glucose oxidase enzyme to the counter electrode may be
decreased. Since the enzyme depletes oxygen locally, the counter
electrode would best be situated at a location distant from the
enzyme. This could be achieved by depleting the enzyme from or
inactivating the enzyme located in the region near and over the
counter electrode by methods known to those skilled in the art such
as laser ablation, or chemical ablation. Alternatively, the
membrane could be covered with an additional domain where glucose
is selectively blocked from the area over the counter
electrode.
[0051] In particular, the present invention reduces the potential
for electrode poisoning by positioning all electrodes underneath a
multi-region membrane so that there is control of the species
reaching the electrode surfaces. These membranes allow passage of
dissolved oxygen to support the counter electrode reactions at
reasonable negative potentials while rejecting larger molecules
which when reduced would coat the surface of the counter electrode
eventually leading to cascade failure. The positioning of the
counter electrode underneath the membrane assures that all currents
are passing through the same conductive media, thereby reducing
voltage losses due to membrane or solution resistance. In addition,
the counter electrode will be able to collect enough species for
the balancing current while minimizing the need to move towards
negative potential extremes.
[0052] Although the description that follows is primarily directed
at glucose monitoring sensor heads, devices and methods for their
use, the sensor heads, devices and methods of the present invention
are not limited to glucose measurement. Rather, the devices and
methods may be applied to detect and quantitate other analytes
present in biological fluids (including, but not limited to, amino
acids and lactate), especially those analytes that are substrates
for oxidase enzymes [see, e.g., U.S. Pat. No. 4,703,756 to Gough et
al., hereby incorporated by reference].
I. Nature of the Foreign Body Capsule
[0053] Devices and probes that are implanted into subcutaneous
tissue will almost always elicit a foreign body capsule (FBC) as
part of the body's response to the introduction of a foreign
material. Therefore, implantation of a glucose sensor results in an
acute inflammatory reaction followed by building of fibrotic
tissue. Ultimately, a mature FBC comprising primarily a vascular
fibrous tissue forms around the device (Shanker and Greisler,
Inflammation and Biomaterials in Greco R S, ed. Implantation
Biology: The Host Response and Biomedical Devices, pp68-80, CRC
Press (1994)).
[0054] In general, the formation of a FBC has precluded the
collection of reliable, continuous information, reportedly because
of poor vascularization (Updike, S. J. et al., "Principles of
Long-term Fully Implanted Sensors with Emphasis on Radiotelemetric
Monitoring of Blood Glucose from inside a Subcutaneous Foreign Body
Capsule (FBC)" in "Biosensors and the Body," D. M. Fraser, ed.,
1997, pp. 117-38, John Wiley and Sons, New York). Thus, those
skilled in the art have previously attempted to minimize FBC
formation by, for example, using a short-lived needle geometry or
sensor coatings to minimize the foreign body.
[0055] In contrast to the prior art, the teachings of the present
invention recognize that FBC formation is the dominant event
surrounding long-term implantation of any sensor and must be
managed to support, rather than hinder or block, sensor
performance. It has been observed that during the early periods
following implantation of an analyte sensing device, particularly a
glucose sensing device, that glucose sensors function well.
However, after a few days to two or more weeks of implantation,
these devices lose their function.
[0056] We have observed that this lack of sensor function is most
likely due to cells (barrier cells) that associate with the outer
surface of the device and physically block the transport of glucose
into the device (i.e. form a barrier cell layer). Increased
vascularization would not be expected to overcome this blockage.
The present invention contemplates the use of particular
biointerface membrane architectures that interfere with barrier
cell layer formation on the membrane's surface. The present
invention also contemplates the use of these membranes with a
variety of implantable devices (e.g. analyte measuring devices
particularly glucose measuring devices).
II. The Sensor Head
[0057] In one embodiment of the sensor head of the invention, the
body is made of a non-conductive material such as ceramic, glass,
or polymer.
[0058] In a preferred embodiment, the sensor head interface region
may include several different layers and/or membranes that cover
and protect the electrodes of an implantable analyte-measuring
device. The characteristics of these layers and/or membranes are
now discussed in more detail. The layers and/or membranes prevent
direct contact of the biological fluid sample with the electrodes,
while permitting selected substances (e.g., analytes) of the fluid
to pass therethrough for reaction in an enzyme rich domain with
subsequent electrochemical reaction of formed products at the
electrodes.
[0059] It is well known in the art that electrode surfaces exposed
to a wide range of biological molecules may suffer poisoning of
catalytic activity and possible corrosion that could result in
failure. However, utilizing the unique multi-region membrane
architectures of the present invention, the active electrochemical
surfaces of the sensor electrodes are preserved, retaining activity
for extended periods of time in vivo. By limiting access to the
electrochemically reactive surface of the electrodes to a small
number of molecular species such as, for example, molecules having
a molecular weight of about 34 Daltons (the molecular weight of
peroxide) or less, only a small subset of the many molecular
species present in biological fluids are permitted to contact the
sensor. Use of such membranes has enabled sustained function of
devices for over one year in vivo. [0060] A. Multi-Region
Membrane
[0061] The multi-region membrane is constructed of two or more
regions. The multi-region membrane may be provided in a number of
different architectures. In one architecture, the multi-region
membrane includes a first region distant from the electrochemically
reactive surfaces, a second region less distant from the
electrochemically reactive surfaces and a third region adjacent to
the electrochemically reactive surfaces. The first region includes
a cell disruptive domain distant from the electrochemically
reactive surfaces and a cell impermeable domain less distant from
the electrochemically reactive surfaces. The second region is a
glucose exclusion domain and the third region includes a resistance
domain distant from the electrochemically reactive surfaces, an
immobilized enzyme domain less distant from the electrochemically
reactive surfaces, an interference domain less distant from the
electrochemically reactive surfaces than the immobilized enzyme
domain and a hydrogel domain adjacent to the electrochemically
reactive surfaces.
[0062] In another architecture, the multi-region membrane includes
a first region distant from the electrochemically reactive surfaces
and a further region less distant from the electrochemically
reactive surfaces. The first region includes a cell disruptive
domain and a cell impermeable domain as described above. The
"further region" includes a resistance domain, immobilized enzyme
domain, interference domain, and hydrogel domain and serves as the
equivalent of the "third region" described above. In certain
embodiments of the sensor head, the multi-region membrane further
includes an oxygen antenna domain. Each of these domains will now
be described in further detail. [0063] i. Cell Disruptive
Domain
[0064] The domain of the multi-region membrane positioned most
distal to the electrochemically reactive surfaces corresponds to
the cell disruptive domain. This domain includes a material that
supports tissue in-growth and may be vascularized. The cell
disruptive domain prevents formation of the barrier cell layer on
the surface of the membrane, which as described above, blocks the
transport of glucose into the sensor device. A useful cell
disruptive domain is described in a U.S. application entitled
"Membrane for use with Implantable Devices" which was filed on the
same day as the present application. The cell disruptive domain may
be composed of an open-cell configuration having cavities and solid
portions. Cells may enter into the cavities, however, they can not
travel through or wholly exist within the solid portions. The
cavities allow most substances to pass through, including, e.g.,
macrophages.
[0065] The open-cell configuration yields a co-continuous solid
domain that contains greater than one cavity in three dimensions
substantially throughout the entirety of the membrane. In addition,
the cavities and cavity interconnections may be formed in layers
having different cavity dimensions.
[0066] A linear line can be used to define a dimension across a
cavity or solid portion the length of which is the distance between
two points lying at the interface of the cavity and solid portion.
In this way, a substantial number of the cavities are not less than
20 microns in the shortest dimension and not more than 1000 microns
in the longest dimension. Preferably, a substantial number of the
cavities are not less than 25 microns in the shortest dimension and
not more than 500 microns in the longest dimension.
[0067] Furthermore, the solid portion has not less than 5 microns
in a substantial number of the shortest dimensions and not more
than 2000 microns in a substantial number of the longest
dimensions. Preferably, the solid portion is not less than 10
microns in a substantial number of the shortest dimensions and not
more than 1000 microns in a substantial number of the longest
dimensions and most preferably, not less than 10 microns in a
substantial number of the shortest dimensions and not more than 400
microns in a substantial number of the longest dimensions.
[0068] The solid portion may be made of polytetrafluoroethylene or
polyethylene-co-tetrafluoroethylene, for example. Preferably, the
solid portion includes polyurethanes or block copolymers and, most
preferably, includes silicone.
[0069] When non-woven fibers are utilized as the solid portion of
the present invention, the non-woven fibers may be greater than 5
microns in the shortest dimension. Preferably, the non-woven fibers
are about 10 microns in the shortest dimension and most preferably,
the non-woven fibers are greater than or equal to 10 microns in the
shortest dimension.
[0070] The non-woven fibers may be constructed of polypropylene
(PP), polyvinylchloride (PVC), polyvinylidene fluoride (PVDF),
polybutylene terephthalate (PBT), polymethylmethacrylate (PMMA),
polyether ether ketone (PEEK), polyurethanes, cellulosic polymers,
polysulfones, and block copolymers thereof including, for example,
di-block, tri-block, alternating, random and graft copolymers
(block copolymers are discussed in U.S. Pat. Nos. 4,803,243 and
4,686,044, hereby incorporated by reference). Preferably, the
non-woven fibers are comprised of polyolefins or polyester or
polycarbonates or polytetrafluoroethylene.
[0071] A subset of the cell disruptive domain is the oxygen antenna
domain. This domain can act as an oxygen reservoir during times of
minimal oxygen need and has the capacity to provide on demand a
higher oxygen gradient to facilitate oxygen transport across the
membrane. This domain may be composed of a material such as
silicone, that has higher oxygen solubility than aqueous media so
that it concentrates oxygen from the biological fluid surrounding
the biointerface membrane. This enhances function in the enzyme
reaction domain and at the counter electrode surface when glucose
conversion to hydrogen peroxide in the enzyme domain consumes
oxygen from the surrounding domains. Thus, this ability of the
oxygen antenna domain to apply a higher flux of oxygen to critical
domains when needed improves overall sensor function. Preferably,
this domain is composed of silicone and has a thickness of about
100 microns.
[0072] The thickness of the cell disruptive domain is usually not
less than about 20 microns and not more than about 2000 microns.
[0073] ii. Cell Impermeable Domain
[0074] The cell impermeable of the first region is positioned less
distal to the electrochemically reactive surfaces than the cell
disruptive domain of the same region. This domain is impermeable to
host cells, such as macrophages. Cell impermeable domains are
described in U.S. Pat. No. 6,001,067, herein incorporated by
reference, and in copending, commonly owned U.S. application
entitled "Membrane for use with Implantable Devices", U.S. Ser. No.
09/916,386, filed on even date herewith. The inflammatory response
that initiates and sustains a FBC is associated with disadvantages
in the practice of sensing analytes. Inflammation is associated
with invasion of inflammatory response cells (e.g. macrophages)
which have the ability to overgrow at the interface and form
barrier cell layers, which may block transport of glucose across
the biointerface membrane. These inflammatory cells may also
biodegrade many artificial biomaterials (some of which were, until
recently, considered nonbiodegradable). When activated by a foreign
body, tissue macrophages degranulate, releasing from their
cytoplasmic myeloperoxidase system hypochlorite (bleach) and other
oxidative species. Hypochlorite and other oxidative species are
known to break down a variety of polymers, including ether based
polyurethanes, by a phenomenon referred to as environmental stress
cracking. Alternatively, polycarbonate based polyurethanes are
believed to be resistant to environmental stress cracking and have
been termed biodurable. In addition, because hypochlorite and other
oxidizing species are short-lived chemical species in vivo,
biodegradation will not occur if macrophages are kept a sufficient
distance from the enzyme active membrane.
[0075] The present invention contemplates the use of cell
impermeable biomaterials of a few microns thickness or more (i.e.,
a cell impermeable domain) in most of its membrane architectures.
This domain of the biointerface membrane is permeable to oxygen and
may or may not be permeable to glucose and is constructed of
biodurable materials (e.g. for period of several years in vivo)
that are impermeable by host cells (e.g. macrophages) such as for
example polymer blends of polycarbonate based polyurethane and
PVP.
[0076] The thickness of the cell impermeable domain is not less
than about 10 microns and not more than about 100 microns. [0077]
iii. Glucose Exclusion Domain
[0078] The glucose exclusion domain includes a thin, hydrophobic
membrane that is non-swellable and blocks diffusion of glucose
while being permeable to oxygen. The glucose exclusion domain
serves to allow analytes and other substances that are to be
measured or utilized by the sensor to pass through, while
preventing passage of other substances. Preferably, the glucose
exclusion domain is constructed of a material such as, for example,
silicone.
[0079] The glucose exclusion domain has a preferred thickness not
less than about 130 microns, more preferably not less than about 5
and not more than about 75 microns and most preferably not less
than 15 microns and not more than about 50 microns. [0080] iv.
Resistance Domain
[0081] In one embodiment of the sensor head the "third region" or
"further region" of the multi-region membrane includes a resistance
domain. When present, the resistance domain is located more distal
to the electrochemically reactive surfaces relative to other
domains in this region. As described in further detail below, the
resistance domain controls the flux of oxygen and glucose to the
underlying enzyme domain. There is a molar excess of glucose
relative to the amount of oxygen in samples of blood. Indeed, for
every free oxygen molecule in extracellular fluid, there are
typically more than 100 glucose molecules present [Updike et al.,
Diabetes Care 5:207-21(1982)]. However, an immobilized enzyme-based
sensor using oxygen (O.sub.2) as cofactor must be supplied with
oxygen in non-rate-limiting excess in order to respond linearly to
changes in glucose concentration, while not responding to changes
in oxygen tension. More specifically, when a glucose-monitoring
reaction is oxygen-limited, linearity is not achieved above minimal
concentrations of glucose. Without a semipermeable membrane over
the enzyme domain, linear response to glucose levels can be
obtained only up to about 40 mg/dL; however, in a clinical setting,
linear response to glucose levels are desirable up to at least
about 500 mg/dL.
[0082] The resistance domain includes a semipermeable membrane that
controls the flux of oxygen and glucose to the underlying enzyme
domain (i.e., limits the flux of glucose), rendering the necessary
supply of oxygen in non-rate-limiting excess. As a result, the
upper limit of linearity of glucose measurement is extended to a
much higher value than that which could be achieved without the
resistance domain. The devices of the present invention contemplate
resistance domains including polymer membranes with
oxygen-to-glucose permeability ratios of approximately 200:1; as a
result, one-dimensional reactant diffusion is adequate to provide
excess oxygen at all reasonable glucose and oxygen concentrations
found in the subcutaneous matrix [Rhodes et al., Anal. Chem.,
66:1520-1529 (1994)].
[0083] In preferred embodiments, the resistance domain is
constructed of a polyurethane urea/polyurethane-block-polyethylene
glycol blend and has a thickness of not more than about 45 microns,
more preferably not less than about 15 microns, and not more than
about 40 microns and, most preferably, not less than about 20
microns, and not more than about 35 microns. [0084] v. Immobilized
Enzyme Domain
[0085] When the resistance domain is combined with the
cell-impermeable domain, it is the immobilized enzyme domain which
corresponds to the outermost domain of the "third region" or
"further region", i.e. it is located more distal to the
electrochemically reactive surfaces as compared to the other
domains in this region. In one embodiment, the enzyme domain
includes glucose oxidase. In addition to glucose oxidase, the
present invention contemplates the use of a domain impregnated with
other oxidases, e.g., galactose oxidase or uricase, for an
enzyme-based electrochemical glucose sensor to perform well, the
sensor's response must neither be limited by enzyme activity nor
cofactor concentration. Because enzymes, including glucose oxidase,
are subject to deactivation as a function of ambient conditions,
this behavior needs to be accounted for in constructing sensors for
long-term use.
[0086] Preferably, the domain is constructed of aqueous dispersions
of colloidal polyurethane polymers including the enzyme.
Preferably, the coating has a thickness of not less than about 2.5
microns and not more than about 12.5 microns, preferably about 6.0
microns. [0087] vi. Interference Domain
[0088] The interference domain in the "third region" or "further
region" is located less distant from the electrochemically reactive
surfaces than the immobilized enzyme domain in this same region. It
includes a thin membrane that can limit diffusion of molecular
weight species greater than 34 kD. The interference domain serves
to allow analytes and other substances that are to be measured by
the electrodes to pass through, while preventing passage of other
substances, including potentially interfering substances. The
interference domain is preferably constructed of a
polyurethane.
[0089] The interference domain has a preferred thickness of not
more than about 5 microns, more preferably not less than about 0.1
microns, and not more than about 5 microns and, most preferably,
not less than about 0.5 microns, and not more than about 3 microns.
[0090] vii. Hydrogel Domain
[0091] The hydrogel domain is located adjacent to the
electrochemically reactive surfaces. To ensure electrochemical
reaction, the hydrogel domain includes a semipermeable coating that
maintains hydrophilicity at the electrode region of the sensor
interface. The hydrogel domain enhances the stability of the
interference domain of the present invention by protecting and
supporting the membrane that makes up the interference domain.
Furthermore, the hydrogel domain assists in stabilizing operation
of the device by overcoming electrode start-up problems and
drifting problems caused by inadequate electrolyte. The buffered
electrolyte solution contained in the hydrogel domain also protects
against pH-mediated damage that may result from the formation of a
large pH gradient between the hydrophobic interference domain and
the electrode (or electrodes) due to the electrochemical activity
of the electrode(s).
[0092] Preferably, the hydrogel domain includes a flexible,
water-swellable, substantially solid gel-like film having a "dry
film" thickness of not less than about 2.5 microns and not more
than about 12.5 microns; preferably, the thickness is about 6.0
microns. "Dry film" thickness refers to the thickness of a cured
film cast from a coating formulation onto the surface of the
membrane by standard coating techniques
[0093] Suitable hydrogel domains are formed of a curable copolymer
of a urethane polymer and a hydrophilic film-forming polymer.
Particularly preferred coatings are formed of a polyurethane
polymer having anionic carboxylate functional groups and non-ionic
hydrophilic polyether segments, which is crosslinked in the present
of polyvinylpyrrolidone and cured at a moderate temperature of
about 50.degree. C. [0094] B. Electrolyte Phase
[0095] The electrolyte phase is a free-fluid phase including a
solution containing at least one compound, usually a soluble
chloride salt, that conducts electric current. The electrolyte
phase flows over the electrodes and is in contact with the hydrogel
domain. The devices of the present invention contemplate the use of
any suitable electrolyte solution, including standard, commercially
available solutions.
[0096] Generally speaking, the electrolyte phase should have the
same or less osmotic pressure than the sample being analyzed. In
preferred embodiments of the present invention, the electrolyte
phase includes normal saline. [0097] C. Membrane Architectures
[0098] Prior art teaches that an enzyme containing membrane that
resides above an amperometric electrochemical sensor can possess
the same architecture throughout the electrode surfaces. However,
the function of converting glucose into hydrogen peroxide by
glucose oxidase may only by necessary above the working electrode.
In fact, it may be beneficial to limit the conversion of glucose
into hydrogen peroxide above the counter electrode. Therefore, the
present invention contemplates a number of membrane architectures
that include a multi-region membrane wherein the regions include at
least one domain.
[0099] Referring now to FIG. 2A, which shows one desired embodiment
of the general architecture of a three region membrane, first
region 33 is permeable to oxygen and glucose and includes a cell
disruptive domain distant from the electrodes and a cell
impermeable domain less distant from the electrodes. The second
region 31 is permeable to oxygen and includes a glucose exclusion
domain and region three 32 includes a resistance domain, distant
from the electrochemically reactive surfaces, an immobilized enzyme
domain less distant from the electrochemically reactive surfaces,
an interference domain less distant from the electrochemically
reactive surfaces than the immobilized enzyme and a hydrogel domain
adjacent to the electrochemically reactive surfaces. The
multi-region membrane is positioned over the sensor interface 30 of
the non-conductive body 10, covering the working electrode 21, the
reference electrode 20 and the counter electrode 22. The electrodes
are brazed to the sensor head and back filled with epoxy 28.
[0100] In FIG. 2B, the glucose exclusion domain has been positioned
over the electrochemically reactive surfaces such that it does not
cover the working electrode 21. To illustrate this, a hole 35 has
been created in the second region 31 and positioned directly above
the working electrode 21. In this way, glucose is blocked from
entering the underlying enzyme membrane above the counter electrode
22 and O.sub.2 is conserved above the counter electrode because it
is not being consumed by the glucose oxidation reaction. The
glucose-blocking domain is made of a material that allows
sufficient O.sub.2 to pass to the counter electrode. The
glucose-blocking domain may be made of a variety of materials such
as silicone or silicone containing copolymers. Preferably, the
glucose-blocking domain is made of silicone.
[0101] In FIG. 2C, the multi-region membrane is shown as being
constructed of two regions: a first region 33 which includes a cell
disruptive domain and a cell impermeable domain; and a further
region 32. Region 32 is defined herein as including an enzyme
immobilized domain, interference domain, and hydrogel domain and
may also include a resistance domain. Region 32 is referred to as
the "third region" in embodiments where the multi-region membrane
includes three regions. In the embodiment shown, a silicone domain
plug 36 positioned over the counter electrode 22 in order to
eliminate the consumption of O.sub.2 above the counter electrode by
the oxidation of glucose with glucose oxidase. The enzyme
immobilized domain can be fabricated as previously described, then
a hole punched into the domain. The silicone domain plug 36 may be
cut to fit the hole, and then adhered into place, for example, with
silicone adhesive (e.g., MED-1511, NuSil, Carpinteria, Calif.).
[0102] In FIG. 2D, the immobilized enzyme domain of the
multi-region membrane can be fabricated such that active enzyme 37
is positioned only above the working electrode 21. In this
architecture, the immobilized enzyme domain may be prepared so that
the glucose oxidase only exists above the working electrode 21.
During the preparation of the multi-region membrane, the
immobilized enzyme domain coating solution can be applied as a
circular region similar to the diameter of the working electrode.
This fabrication can be accomplished in a variety of ways such as
screen printing or pad printing. Preferably, the enzyme domain is
pad printed during the enzyme membrane fabrication with equipment
as available from Pad Print Machinery of Vermont (Manchester, Vt.).
These architectures eliminate the consumption of O.sub.2 above the
counter electrode 22 by the oxidation of glucose with glucose
oxidase.
[0103] In FIG. 2E, the immobilized enzyme of the multi-region
membrane in region 32 may be deactivated 38 except for the area
covering the working electrode 21. In some of the previous membrane
architectures, the glucose oxidase is distributed homogeneously
throughout the immobilized enzyme domain. However, the active
enzyme need only reside above the working electrode. Therefore, the
enzyme may be deactivated 38 above the counter 22 and reference 20
electrodes by irradiation. A mask that covers the working electrode
21, such as those used for photolithography can be placed above the
membrane. In this way, exposure of the masked membrane to
ultraviolet light deactivates the glucose oxidase in all regions
except that covered by the mask.
[0104] FIG. 2F shows an architecture in which the third region 32
which includes immobilized enzyme only resides over the working
electrode 21. In this architecture, consumption of O.sub.2 above
the counter electrode 22 by the oxidation of glucose with glucose
oxidase is eliminated. [0105] D. The Electrode Assembly
[0106] The electrode assembly of this invention comprises a
non-conductive body and three electrodes affixed within the body
having electrochemically reactive surfaces at one location on the
body and an electronic connection means at another location on the
body and may be used in the manner commonly employed in the making
of amperometric measurements. A sample of the fluid being analyzed
is placed in contact with a reference electrode, e.g.,
silver/silver-chloride, a working electrode which is preferably
formed of platinum, and a counter electrode which is preferably
formed of gold or platinum. The electrodes are connected to a
galvanometer or polarographic instrument and the current is read or
recorded upon application of the desired D.C. bias voltage between
the electrodes.
[0107] The ability of the present device electrode assembly to
accurately measure the concentration of substances such as glucose
over a broad range of concentrations in fluids including undiluted
whole blood samples enables the rapid and accurate determination of
the concentration of those substances. That information can be
employed in the study and control of metabolic disorders including
diabetes.
[0108] The present invention contemplates several structural
architectures that effectively increase the electrochemically
reactive surface of the counter electrode. In one embodiment, the
diameter of wire used to create the counter electrode is at least
twice the diameter of the working electrode. In this architecture,
it is preferable that the electrochemically reactive surface of the
counter electrode be not less than about 2 and not more than about
100 times the surface area of the working electrode. More
preferably, the electrochemically reactive surface of the counter
electrode is not less than about 2 and not more than about 50, not
less than about 2 and not more than about 25 or not less than about
2 and not more than about 10 times the surface area of the working
electrode. In another embodiment, the electrochemically reactive
surface is larger that the wire connecting this surface to the
sensor head. In this architecture, the electrode could have a
cross-sectional view that resembles a "T". The present invention
contemplates a variety of configurations of the electrode head that
would provide a large reactive surface, while maintaining a
relatively narrow connecting wire. Such configurations could be
prepared by micromachining with techniques such as reactive ion
etching, wet chemical etching and focused ion beam machining as
available from Norsam Technologies (Santa Fe, N. Mex.).
[0109] In another embodiment, the diameter of the counter electrode
is substantially similar to the working electrode; however, the
surface of the counter electrode has been modified to increase the
surface area such that it has at least twice the surface area of
the working electrode. More specifically the counter electrodes
surface may be textured, effectively increasing its surface area
without significantly increasing its diameter. This may be
accomplished by a variety of methods known to those skilled in the
art including, such as acid etching. The electrochemically reactive
surface may be provided in a variety of shapes and sizes (e.g.
round, triangular, square or free form) provided that it is at
least twice the surface area of the working electrode.
[0110] In all of the architectures described, the electrodes are
prepared from a 0.020'' diameter wire having the desired modified
reactive surface. The electrodes are secured inside the
non-conductive body by brazing. The counter electrode is preferably
made of gold or platinum.
III. Analyte Measuring Device
[0111] A preferred embodiment of an analyte measuring device
including a sensor head according to the present invention is shown
in FIG. 6A. In this embodiment, a ceramic body 1 and ceramic head
10 houses the sensor electronics that include a circuit board 2, a
microprocessor 3, a battery 4, and an antenna 5. Furthermore, the
ceramic body 1 and head 10 possess a matching taper joint 6 that is
sealed with epoxy. The electrodes are subsequently connected to the
circuit board via a socket 8.
[0112] As indicated in detail in FIG. 6B, three electrodes protrude
through the ceramic head 10, a platinum working electrode 21, a
platinum counter electrode 22 and a silver/silver chloride
reference electrode 20. Each of these is hermetically brazed 26 to
the ceramic head 10 and further secured with epoxy 28. The sensing
region 24 is covered with a multi-region membrane described above
and the ceramic head 10 contains a groove 29 so that the membrane
may be affixed into place with an o-ring.
IV. Experimental
[0113] The following examples serve to illustrate certain preferred
embodiments and aspects of the present invention and are not to be
construed as limiting the scope thereof.
[0114] In the preceding description and the experimental disclosure
which follows, the following abbreviations apply: Eq and Eqs
(equivalents); mEq (milliequivalents); M (molar); mM (millimolar)
.mu.M (micromolar); N (Normal); mol (moles); mmol (millimoles);
.mu.mol (micromoles); nmol (nanomoles); g (grams); mg (milligrams);
.mu.g (micrograms); Kg (kilograms); L (liters); mL (milliliters);
dL (deciliters); .mu.L (microliters); cm (centimeters); mm
(millimeters); .mu.m (micrometers); nm (nanometers); h and hr
(hours); min. (minutes); s and sec. (seconds); .degree. C. (degrees
Centigrade); Astor Wax (Titusville, Pa.); BASF Wyandotte
Corporation (Parsippany, N.J.); Data Sciences, Inc. (St. Paul,
Minn.); DuPont (DuPont Co., Wilmington, Del.); Exxon Chemical
(Houston, Tex.); GAF Corporation (New York, N.Y.); Markwell Medical
(Racine, Wis.); Meadox Medical, Inc. (Oakland, N.J.); Mobay (Mobay
Corporation, Pittsburgh, Pa.); NuSil Technologies (Carpenteria,
Calif.) Sandoz (East Hanover, N.J.); and Union Carbide (Union
Carbide Corporation; Chicago, Ill.).
EXAMPLE 1
Preparation of the Multi-Region Membrane
A. Preparation of the First Region
[0115] The cell disruptive domain may be an ePTFE filtration
membrane and the cell impermeable domain may then be coated on this
domain layer. The cell impermeable domain was prepared by placing
approximately 706 gm of dimethylacetamide (DMAC) into a 3 L
stainless steel bowl to which a polycarbonateurethane solution
(1325 g, Chronoflex AR 25% solids in DMAC and 5100 cp) and
polyvinylpyrrolidone (125 g, Plasdone K-90 D) are added. The bowl
was then fitted to a planetary mixer with a paddle type blade and
the contents were stirred for 1 hour at room temperature. This
solution was then coated on the cell disruptive domain by knife
edge drawn at a gap of 0.006'' and dried at 60.degree. C. for 24
hours.
[0116] Alternatively, the polyurethane polyvinylpyrrolidone
solution prepared above can be coated onto a PET release liner
using a knife over roll coating machine. This material is then
dried at 305.degree. F. for approximately 2 minutes. Next the ePTFE
membrane is immersed in 50:50 (w/v) mixture of THF/DMAC and then
placed atop the coated polyurethane polyvinylpyrrolidone material.
Light pressure atop the assembly intimately embeds the ePTFE into
the polyurethane polyvinylpyrrolidone. The membrane is then dried
at 60.degree. C. for 24 hours.
B. Preparation of the Glucose Exclusion Domain
[0117] An oxime cured silicone dispersion (NuSil Technologies,
MED-6607) was cast onto a polypropylene sheet and cured at
40.degree. C. for three days.
C. Preparation of the Third Region
[0118] The "third region" or "further region" includes a resistance
domain, an immobilized enzyme domain, an interference domain and a
hydrogel domain. The resistance domain was prepared by placing
approximately 281 gm of dimethylacetamide into a 3 L stainless
steel bowl to which a solution of polyetherurethaneurea (344 gm of
Chronothane H, 29,750 cp at 25% solids in DMAC). To this mixture
was added another polyetherurethaneurea (312 gm, Chronothane 1020,
6275 cp at 25% solids in DMAC.) The bowl was fitted to a planetary
mixer with a paddle type blade and the contents were stirred for 30
minutes at room temperature. The resistance domain coating
solutions produced is coated onto a PET release liner (Douglas
Hansen Co., Inc. Minneapolis, Minn.) using a knife over roll set at
a 0.012'' gap. This film is then dried at 305.degree. F. The final
film is approximately 0.0015'' thick.
[0119] The immobilized enzyme domain was prepared by placing 304 gm
polyurethane latex (Bayhydrol 140 AQ, Bayer, Pittsburgh, Pa.) into
a 3 L stainless steel bowl to which 51 gm of pyrogen free water and
5.85 gm of glucose oxidase (Sigma type VII from Aspergillus niger)
is added. The bowl was then fitted to a planetary mixer with a
whisk type blade and the mixture was stirred for 15 minutes.
Approximately 24 hr prior to coating a solution of glutaraldehyde
(15.4 mL of a 2.5% solution in pyrogen free water) and 14 mL of
pyrogen free water was added to the mixture. The solution was mixed
by inverting a capped glass bottle by hand for about 3 minutes at
room temperature. This mixture was then coated over the resistance
domain with a #10 Mayer rod and dried above room temperature
preferably at about 50.degree. C.
[0120] The interference domain was prepared by placing 187 gm of
tetrahydrofuran into a 500 mL glass bottle to which an 18.7 gm
aliphatic polyetherurethane (Tecoflex SG-85A, Thermedics Inc.,
Woburn, Mass.) was added. The bottle was placed onto a roller at
approximately 3 rpm within an oven set at 37.degree. C. The mixture
was allowed to roll for 24 hr. This mixture was coated over the
dried enzyme domain using a flexible knife and dried above room
temperature preferably at about 50.degree. C.
[0121] The hydrogel domain was prepared by placing 388 gm of
polyurethane latex (Bayhydrol 123, Bayer, Pittsburgh, Pa. in a 3 L
stainless steel bowl to which 125 gm of pyrogen free water and 12.5
gm polyvinylpyrrolidone (Plasdone K-90D) was added. The bowl was
then fitted to a planetary mixer with a paddle type blade and
stirred for 1 hr at room temperature. Within 30 minutes of coating
approximately 13.1 mL of carbodiimide (UCARLNK) was added and the
solution was mixed by inverting a capped polyethylene jar by hand
for about 3 min at room temperature. This mixture was coated over
the dried interference domain with a #10 Mayer rod and dried above
room temperature preferably at about 50.degree. C.
[0122] In order to affix this multi-region membrane to a sensor
head, it is first placed into buffer for about 2 minutes. It is
then stretched over the nonconductive body of sensor head and
affixed into place with an o-ring.
EXAMPLE 2
In Vitro Evaluation of Sensor Devices
[0123] This example describes experiments directed at sensor
function of several sensor devices contemplated by the present
invention.
[0124] In vitro testing of the sensor devices was accomplished in a
manner similar to that previously described. [Gilligan et al.,
Diabetes Care 17:882-887 (1994)]. Briefly, devices were powered on
and placed into a polyethylene container containing phosphate
buffer (450 ml, pH 7.30) at 37.degree. C. The container was placed
onto a shaker (Lab Line Rotator, model 1314) set to speed 2. The
sensors were allowed to equilibrate for at least 30 minutes and
their output value recorded. After this time, a glucose solution
(9.2 ml of 100 mg/ml glucose in buffer) was added in order to raise
the glucose concentration to 200 mg/dl within the container. The
sensors were allowed to equilibrate for at least 30 minutes and
their output value recorded. Again, a glucose solution (9.4 ml of
100 mg/ml glucose in buffer) was added in order to raise the
glucose concentration to 400 mg/dl within the container. The
sensors were allowed to equilibrate for at least 30 minutes and
their output value recorded. In this way, the sensitivity of the
sensor to glucose is given as the slope of sensor output versus
glucose concentration. The container was then fitted with an
O.sub.2 meter (WTW, model Oxi-340) and a gas purge. A mixture of
compressed air and nitrogen was used to decrease the O.sub.2
concentration. Sensor output was recorded at an ambient O.sub.2
level, then sensor output was recorded for the following O.sub.2
concentrations; 1 mg/L, 0.85 to 0.75 mg/L, 0.65 to 0.55 mg/L and
0.40 to 0.30 mg/L. In this way, the function of the sensor could be
compared to its function at ambient O.sub.2.
[0125] Sensor devices like the one shown in FIGS. 6A and 6B, which
included inventive sensor heads having a multi-region membrane with
the architecture shown in FIG. 2B, were tested in vitro. Eight of
these devices were fitted with membranes that possessed a 0.020''
diameter hole, four with a 0.0015'' thick polyurethane (Chronoflex
AR, CardioTech International Inc.) and four with a 0.032'' thick
silicone (MED-1511, NuSil Technologies Inc.). The hole was
positioned above the working electrode and both membranes were
secured to the device with an o-ring. Four control devices were
also tested which were fitted with a multi-region membrane which
lacked region 31 shown in FIB. 2B.
[0126] As discussed above, for oxygen to be consumed in the sensing
region 32 above the electrodes, glucose is required. By placing
region 31 shown in FIG. 2B, which includes a glucose blocking
domain, above all areas other than above the working electrode 21,
oxygen consumption in areas other than working electrode areas is
limited. In contrast, by eliminating region 31 in the control
devices, less overall oxygen becomes available to electrode
surfaces due to the increased availability of glucose.
[0127] The devices were activated, placed into a 500
ml-polyethylene container with sodium phosphate buffered solution
(300 ml, pH 7.3) and allowed to equilibrate. Each device's baseline
value was recorded. Then 12 ml of glucose solution (100 mg/ml in
sodium phosphate buffer) was added to the container so that the
total glucose concentration became 400 mg/dL. After this, the
container was covered and fitted with an oxygen sensor and a source
of nitrogen and compressed air. In this way, the oxygen
concentration was controlled with a gas sparge. A glucose value was
recorded for each device at decreasing oxygen concentrations from
ambient to approximately 0.1 mg/L.
[0128] FIG. 7 graphically represents the formation of a device of
the present invention utilizing the multi-region membrane
architecture in FIG. 2B in vitro. The data is expressed in percent
Device Function at 400 mg/dL glucose vs. oxygen concentration. The
percent function of the device is simply the device output at any
given oxygen concentration divided by that device's output at
ambient oxygen. The results from FIG. 7 indicate that inventive
sensor devices containing the silicone membrane have better
function at lower oxygen concentrations relative to both the
control devices and the devices containing the polyurethane
membrane. For example, at an oxygen concentration of about 0.5
mg/L, devices containing the silicone membrane are providing 100%
output as compared to 80% output for the control devices.
EXAMPLE 3
The Effect of Varying the Size and Material of the Counter
Electrode on Sensor Response and Accuracy
[0129] An in vitro testing procedure used in this example was
similar to that described in Example 2. Six devices similar to the
one shown in FIG. 6A and 6B were fitted with the multi-region
membrane described herein. Two of these tested devices were
comparative devices that possessed Pt counter electrodes having a
0.020'' diameter; this diameter provided for an electrochemically
reactive surface of the counter electrode which was substantially
equal to the surface area of the working electrode, as
schematically shown in FIG. 8. In FIG. 8, the electrode-membrane
region includes two distinct regions, the compositions and
functions of which have already been described. Region 32 includes
an immobilized enzyme. Region 33 includes a cell disruptive domain
and a cell impermeable domain. The top ends of electrodes 21
(working), 20 (reference) and 22 (counter) are in contact with an
electrolyte phase 30, a free-flowing phase. Two other tested
devices possessed Pt counter electrodes having a 0.060'' diameter.
Finally, two additional devices possessed Au counter electrodes
having a 0.060'' diameter. The 0.006'' diameter devices provided
for an electrochemically reactive surface of the counter electrode
which was approximately six times the surface area of the working
electrode. Each of the devices including counter electrodes of
0.060'' diameter include a multi-region membrane above the
electrode region which is similar to that shown in FIG. 8.
[0130] The devices were activated, placed into a 500
ml-polyethylene container with sodium phosphate buffered solution
(300 ml, pH 7.3) and allowed to equilibrate. Each device's baseline
value was recorded. Then 12 ml of glucose solution (100 mg/ml in
sodium phosphate buffer) was added to the container so that the
total glucose concentration became 400 mg/dL. After this, the
container was covered and fitted with an oxygen sensor and a source
of nitrogen and compressed air. In this way, the oxygen
concentration was controlled with a gas sparge. A counter electrode
voltage was recorded for each device at decreasing oxygen
concentrations from ambient to approximately 0.1 mg/L.
[0131] FIG. 9 graphically presents the counter electrode voltage as
a function of oxygen concentration and 400 mg/dL glucose. This
figure demonstrates that both the large Pt and Au counter electrode
devices do not begin to reach the circuitry limits at low oxygen
concentrations. Therefore, increased performance and accuracy can
be obtained from a counter electrode that has an electrochemical
reactive surface greater than the surface area of the working
electrode.
[0132] The description and experimental materials presented above
are intended to be illustrative of the present invention while not
limiting the scope thereof. It will be apparent to those skilled in
the art that variations and modifications can be made without
departing from the spirit and scope of the present invention.
* * * * *