U.S. patent application number 17/012410 was filed with the patent office on 2021-04-29 for systems for removing air from the fluid circuits of a plasma processing system.
The applicant listed for this patent is HDL Therapeutics, Inc.. Invention is credited to Hollis Bryan Brewer, JR., Michael M. Matin, Timothy Jon Perlman.
Application Number | 20210121612 17/012410 |
Document ID | / |
Family ID | 1000005324422 |
Filed Date | 2021-04-29 |
![](/patent/app/20210121612/US20210121612A1-20210429\US20210121612A1-2021042)
United States Patent
Application |
20210121612 |
Kind Code |
A1 |
Brewer, JR.; Hollis Bryan ;
et al. |
April 29, 2021 |
Systems for Removing Air from the Fluid Circuits of a Plasma
Processing System
Abstract
The present specification discloses plasma processing systems
that include a number of different fluid flow circuits that are
defined by sources of fluid, fluid lines, fluid flow paths, waste
containers, a mixer, a separator, valves, and a pump. The systems
also include a connector tube and a solvent extraction device,
wherein the connector tube and solvent extraction device are
configured to be alternatively inserted in a same position along a
fluid flow line. In addition, the systems include a controller that
is configured to execute a plurality of programmatic instructions
to open and close each of a first fluid flow line valve, a second
fluid flow line valve, a third fluid flow line valve, and a fourth
fluid flow line valve in a predetermined sequence.
Inventors: |
Brewer, JR.; Hollis Bryan;
(Potomac, MD) ; Matin; Michael M.; (Short Hills,
NJ) ; Perlman; Timothy Jon; (Pleasanton, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
HDL Therapeutics, Inc. |
Vero Beach |
FL |
US |
|
|
Family ID: |
1000005324422 |
Appl. No.: |
17/012410 |
Filed: |
September 4, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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16198672 |
Nov 21, 2018 |
|
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17012410 |
|
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62589919 |
Nov 22, 2017 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/575 20130101;
B01D 11/0446 20130101; B01D 11/0492 20130101; A61M 1/0281 20130101;
B01D 15/10 20130101; A61M 1/0209 20130101; A61K 35/16 20130101 |
International
Class: |
A61M 1/02 20060101
A61M001/02; A61K 31/575 20060101 A61K031/575; A61K 35/16 20060101
A61K035/16; B01D 11/04 20060101 B01D011/04; B01D 15/10 20060101
B01D015/10 |
Claims
1. A plasma processing system comprising: a source of saline; a
first fluid flow line; a first fluid flow line valve positioned
between the source of saline and the first fluid flow line; a
second fluid flow line; a second fluid flow line valve positioned
between the first fluid flow line and a second fluid flow line; a
third fluid flow line; a pump positioned between the second fluid
flow line and the third fluid flow line; a prime waste container in
fluid communication with the third fluid flow line; a fourth fluid
flow line; a fourth fluid flow line valve positioned between the
first fluid flow line and the fourth fluid flow line; a third fluid
flow path valve positioned along the third fluid flow path; a
connector tube; a solvent extraction device, wherein the connector
tube and solvent extraction device are configured to be
alternatively inserted in a same position along the second fluid
flow line; and a controller, wherein the controller is configured
to execute a plurality of programmatic instructions to open and
close each of the first fluid flow line valve, second fluid flow
line valve, third fluid flow line valve, and fourth fluid flow line
valve in a predetermined sequence.
2. The plasma processing system of claim 1, wherein the solvent
extraction device is a charcoal column.
3. The plasma processing system of claim 1, wherein the first fluid
flow path is not in fluid communication with a source of plasma or
a source of solvent.
4. The plasma processing system of claim 1, further comprising a
separator, wherein the fourth fluid flow line is in fluid
communication with the separator.
5. The plasma processing system of claim 1, further comprising an
output plasma container, wherein the output plasma container is in
fluid communication with the pump.
6. The plasma processing system of claim 1, wherein the controller
is configured to execute a plurality of programmatic instructions
to open and close each of the first fluid flow line valve, second
fluid flow line valve, third fluid flow line valve, and fourth
fluid flow line valve such that the first fluid flow line, second
fluid flow line, and fourth fluid flow line are filled with saline
before the fourth fluid flow line is closed.
7. The plasma processing system of claim 1, wherein the controller
is configured to execute a plurality of programmatic instructions
to open and close each of the first fluid flow line valve, second
fluid flow line valve, third fluid flow line valve, and fourth
fluid flow line valve such that the first fluid flow line, second
fluid flow line, and fourth fluid flow line are filled with saline
before the third fluid flow line is opened.
8. The plasma processing system of claim 1, wherein the controller
is configured to execute a plurality of programmatic instructions
to activate the pump and open and close each of the first fluid
flow line valve, second fluid flow line valve, third fluid flow
line valve, and fourth fluid flow line valve such that fluid in the
first fluid flow line and second fluid flow line is directed to the
prime waste container.
9. A plasma processing system comprising: a source of saline; a
plurality of fluid flow lines, wherein the plurality of fluid flow
lines comprise a first fluid flow line, a second fluid flow line, a
third fluid flow line, and a fourth fluid flow line; a plurality of
fluid flow line valves positioned along each of the plurality of
fluid flow lines; a connector tube; a solvent extraction device,
wherein the connector tube and solvent extraction device are
configured to be alternatively inserted in a same position along
the second fluid flow line; and a controller, wherein the
controller is configured to execute a plurality of programmatic
instructions to open and close each of the plurality of fluid flow
line valves in a predetermined sequence.
10. The plasma processing system of claim 9, wherein the solvent
extraction device is a charcoal column.
11. The plasma processing system of claim 9, further comprising a
separator, wherein the fourth fluid flow line is in fluid
communication with the separator.
12. The plasma processing system of claim 9, wherein the plurality
of fluid flow line valves comprise a first fluid flow line valve
positioned between the source of saline and the first fluid flow
line, a second fluid flow line valve positioned between the first
fluid flow line and the second fluid flow line, a fourth fluid flow
line valve positioned between the first fluid flow line and the
fourth fluid flow line, and a third fluid flow path valve
positioned along the third fluid flow path.
13. The plasma processing system of claim 12, further comprising a
pump positioned between the second fluid flow line and the third
fluid flow line.
14. The plasma processing system of claim 13, further comprising a
prime waste container in fluid communication with the third fluid
flow line.
15. The plasma processing system of claim 12, wherein the
controller is configured to execute a plurality of programmatic
instructions to open and close each of the first fluid flow line
valve, second fluid flow line valve, third fluid flow line valve,
and fourth fluid flow line valve such that the first fluid flow
line, second fluid flow line, and fourth fluid flow line are filled
with saline before the fourth fluid flow line is closed.
16. The plasma processing system of claim 12, wherein the
controller is configured to execute a plurality of programmatic
instructions to open and close each of the first fluid flow line
valve, second fluid flow line valve, third fluid flow line valve,
and fourth fluid flow line valve such that the first fluid flow
line, second fluid flow line, and fourth fluid flow line are filled
with saline before the third fluid flow line is opened.
17. The plasma processing system of claim 14, wherein the
controller is configured to execute a plurality of programmatic
instructions to activate the pump and open and close each of the
first fluid flow line valve, second fluid flow line valve, third
fluid flow line valve, and fourth fluid flow line valve such that
fluid in the first fluid flow line and second fluid flow line is
directed to the prime waste container.
Description
CROSS-REFERENCE
[0001] The present application is a continuation of U.S. patent
application Ser. No. 16/198,672, entitled "Systems and Methods for
Priming Fluid Circuits of a Plasma Processing System" and filed on
Nov. 21, 2018, which, in turn, relies on U.S. Provisional Patent
Application Ser. No. 62/589,919, entitled "Systems and Methods for
Causing Regression of Arterial Plaque" and filed on Nov. 22, 2017,
for priority. The above-mentioned applications are herein
incorporated by reference in their entirety.
FIELD
[0002] The present invention generally relates to systems,
apparatus and methods for removing lipids from HDL particles while
leaving LDL particles substantially intact, via the extracorporeal
treatment of blood plasma using either a single solvent or multiple
solvents, in order to regress vulnerable arterial plaques, which is
implicated in many disease states. More specifically, the presently
disclosed inventions address the priming of the plasma processing
system and the management of waste, particularly solvent waste,
generated by the described treatment processes.
BACKGROUND
[0003] Familial Hypercholesterolemia (FH) is an inherited genetic
autosomal dominant disease characterized by markedly elevated low
density lipoprotein (LDL), tendon xanthomas, and premature coronary
heart disease, caused by mutations of "FH genes," which include the
LDL-receptor (LDLR), apolipoprotein B-100 (APOB) or proprotein
convertase subtilisin/kexin type 9 (PCSK9). FH produces a
clinically recognizable pattern that consists of severe
hypercholesterolemia due to the accumulation of LDL in the plasma,
cholesterol deposition in tendons and skin, as well as a high risk
of atherosclerosis manifesting almost exclusively as coronary
artery disease (CAD). In FH patients, this genetic mutation makes
the liver unable to effectively metabolize (or remove) excess
plasma LDL, resulting in increased LDL levels.
[0004] If an individual has inherited a defective FH gene from one
parent, the form of FH is called Heterozygous FH. Heterozygous FH
is a common genetic disorder, inherited in an autosomal dominant
pattern, occurring in approximately 1:500 people in most countries.
If the individual has inherited a defective FH gene from both
parents, the form of FH is called Homozygous FH. Homozygous FH is
very rare, occurring in about 1 in 160,000 to one million people
worldwide, and results in LDL levels>700 mg/dl, 10 fold higher
than the ideal 70 mg/dl level desired for patients with CVD. Due to
the high LDL levels, patients with Homozygous FH have aggressive
atherosclerosis (narrowing and blocking of blood vessels) and early
heart attacks. This process starts before birth and progresses
rapidly. It can affect the coronary arteries, carotid arteries,
aorta, and aortic valve.
[0005] Heterozygous FH (HeFH) is normally treated with statins,
bile acid sequestrants, or other lipid lowering agents that lower
cholesterol levels, and/or by offering genetic counseling.
Homozygous FH (HoFH) often does not respond adequately to medical
therapy and may require other treatments, including LDL apheresis
(removal of LDL in a method similar to dialysis), ileal bypass
surgery to dramatically lower their LDL levels, and occasionally
liver transplantation. A few medications have recently been
approved for use by HoFH subjects. However, these medications lower
LDL only, and modestly contribute to slowing, but not stopping,
further progression of atherosclerosis. Additionally, these
medications are known to have significant side-effects.
[0006] Cholesterol is synthesized by the liver or obtained from
dietary sources. LDL is responsible for transferring cholesterol
from the liver to tissues at different sites in the body. However,
if LDL collects on the arterial walls, it undergoes oxidation
caused by oxygen free radicals liberated from the body's chemical
processes and interacts deleteriously with the blood vessels. The
modified LDL causes white blood cells in the immune system to
gather at the arterial walls, forming a fatty substance called
plaque and injuring cellular layers that line blood vessels. The
modified oxidized LDL also reduces the level of nitric oxide, which
is responsible for relaxing the blood vessels and thereby allowing
the blood to flow freely. As this process continues, the arterial
walls slowly constrict, resulting in hardening of the arteries and
thereby reducing blood flow. The gradual build-up of plaque can
result in blockage of a coronary vessel and ultimately in a heart
attack. The plaque build up can also occur in peripheral vessels
such as the legs and this condition is known as peripheral arterial
disease.
[0007] Obstructions can also appear in blood vessels that supply
blood to the brain, which can result in ischemic strokes. The
underlying condition for this type of obstruction is the
development of fatty deposits lining the vessel walls. It is known
that at least 2.7% of men and women over the age of 18 in the
United States have a history of stroke. Prevalence of stroke is
also known to be higher with increasing age. With the increase in
the aging population, the prevalence of stroke survivors is
projected to increase, especially among elderly women. A
considerable portion of all strokes (at least 87%) are ischemic in
nature.
[0008] Further, it has been shown that hypercholesterolemia and
inflammation are two dominant mechanisms implicated in the
development of atherosclerosis. There is significant overlap
between vascular risk factors for both Alzheimer's disease and
atherosclerosis. Inflammation has been implicated in Alzheimer's
disease pathogenesis and it is suggested that abnormalities in
cholesterol homeostasis may have a role as well. In addition, many
of the contributory factors in atherogenesis also contribute to
Alzheimer's disease. Specifically, in cell cultures, increased and
decreased cholesterol levels promote and inhibit the formation of
beta amyloid (A.beta.) from Amyloid Precursor Protein (APP),
respectively. Thus, the use of treatments with proven effects on
the process of atherosclerosis may be one method for treating the
progression of the Alzheimer's disease.
[0009] Another common cardiovascular disease that occurs due to
development of atherosclerosis (hardening and narrowing of the
arteries) within the elastic lining inside a coronary artery, is
Coronary Artery Disease (CAD), also known as Ischemic Heart Disease
(IHD). On the basis of a statistical data collected from 2009 to
2012, an estimated 15.5 million Americans.gtoreq.20 years of age
have CAD. The total CAD prevalence in the United States is 6.2% of
adults.gtoreq.20 years of age.
[0010] An acute decrease in blood flow in the coronary arteries may
result in part of the heart muscle unable to function properly.
This condition is known as Acute Coronary Syndrome (ACS). A
conservative estimate for the number of hospital discharges with
ACS in 2010 is 625,000.
[0011] In contrast to LDL, high plasma HDL levels are desirable
because they play a major role in "reverse cholesterol transport",
where the excess cholesterol is transferred from tissue sites to
the liver where it is eliminated. Optimal total cholesterol levels
are 200 mg/dl or below with a LDL cholesterol level of 160 mg/dl or
below and a HDL-cholesterol level of 45 mg/dl for men and 50 mg/dl
for women. Lower LDL levels are recommended for individuals with a
history of elevated cholesterol, atherosclerosis or coronary artery
disease. High levels of LDL increase the lipid content in coronary
arteries resulting in formation of lipid filled plaques that are
vulnerable to rupture. On the other hand, HDL has been shown to
decrease the lipid content in the lipid filled plaques, reducing
the probability of rupture. In the last several years, clinical
trials of low density lipoprotein (LDL)-lowering drugs have
definitively established that reductions in LDL are associated with
a 30-45% decrease in clinical cardiovascular disease (CVD) events.
CVD events include events occurring in diseases such as HoFH, HeFH,
and peripheral arterial disease. Despite lowered LDL, however, many
patients continue to have cardiac events. Low levels of HDL are
often present in high risk subjects with CVD, and epidemiological
studies have identified HDL as an independent risk factor that
modulates CVD risk. In addition to epidemiologic studies, other
evidence suggests that raising HDL would reduce the risk of CVD.
There has been increasing interest in changing plasma HDL levels by
dietary, pharmacological or genetic manipulations as a potential
strategy for the treatment of CVD including HoFH, HeFH, Ischemic
stroke, CAD, ACS, and peripheral arterial disease and for treating
the progression of Alzheimer's Disease.
[0012] The protein component of LDL, known as apolipoprotein-B
(ApoB), and its products, comprise atherogenic elements. Elevated
plasma LDL levels and reduced HDL levels are recognized as primary
causes of coronary disease. ApoB is in highest concentration in LDL
particles and is not present in HDL particles. Apolipoprotein A-I
(ApoA-I) and apolipoprotein A-II (ApoA-II) are found in HDL. Other
apolipoproteins, such as ApoC and its subtypes (C-I, C-II and
C-III), ApoD, and ApoE are also found in HDL. ApoC and ApoE are
also observed in LDL particles.
[0013] Numerous major classes of HDL particles including HDL2b,
HDL2a, HDL3a, HDL3b and HDL3 have been reported. Various forms of
HDL particles have been described on the basis of electrophoretic
mobility on agarose as two major populations, a major fraction with
.alpha.-HDL mobility and a minor fraction with migration similar to
VLDL. This latter fraction has been called pre-.beta. HDL and these
particles are the most efficient HDL particle subclass for inducing
cellular cholesterol efflux.
[0014] The HDL lipoprotein particles are comprised of ApoA-I,
phospholipids and cholesterol. The pre-.beta. HDL particles are
considered to be the first acceptors of cellular free cholesterol
and are essential in eventually transferring free and esterified
cholesterol to .alpha.-HDL. Pre-.beta. HDL particles may transfer
cholesterol to .alpha.-HDL or be converted to .alpha.-HDL. The
alpha HDL transfers cholesterol to the liver, where excess
cholesterol can be removed from the body.
[0015] HDL levels are inversely correlated with atherosclerosis and
coronary artery disease. Once cholesterol-carrying .alpha.-HDL
reaches the liver, the .alpha.-HDL particles divest of the
cholesterol and transfer the free cholesterol to the liver. The
.alpha.-HDL particles (divested of cholesterol) are subsequently
converted to pre-.beta. HDL particles and exit the liver, which
then serve to pick up additional cholesterol within the body and
are converted back to .alpha.-HDL, thus repeating the cycle.
Accordingly, what is needed is a method to decrease or remove
cholesterol from these various HDL particles, especially the
.alpha.-HDL particles, so that they are available to remove
additional cholesterol from cells.
[0016] Hyperlipidemia (or abnormally high concentration of lipids
in the blood) may be treated by changing a patient's diet. However,
diet as a primary mode of therapy requires a major effort on the
part of patients, physicians, nutritionists, dietitians, and other
health care professionals and thus undesirably taxes the resources
of health professionals. Another negative aspect of this therapy is
that its success does not rest exclusively on diet. Rather, success
of dietary therapy depends upon a combination of social,
psychological, economic, and behavioral factors. Thus, therapy
based only on correcting flaws within a patient's diet, is not
always successful.
[0017] In instances when dietary modification has been
unsuccessful, drug therapy has been used as adjunctive therapy.
Such therapy has included use of commercially available
hypolipidemic drugs administered alone or in combination with other
therapies as a supplement to dietary control. These drugs, called
statins, include lovastatin, pravastatin, simvastatin, fluvastatin,
atorvastatin, and cerivastatin. Statins are particularly effective
for lowering LDL levels and are also effective in the reduction of
triglycerides, apparently in direct proportion to their
LDL-lowering effects. Statins raise HDL levels, but to a lesser
extent than other anti-cholesterol drugs. Statins also increase
nitric oxide, which, as described above, is reduced in the presence
of oxidized LDL.
[0018] Bile acid resins, another drug therapy, work by binding with
bile acid, a substance made by the liver using cholesterol as one
of the primary manufacturing components. Because the drugs bind
with bile acids in the digestive tract, they are then excreted with
the feces rather than being absorbed into the body. The liver, as a
result, must take more cholesterol from the circulation to continue
constructing bile acids, resulting in an overall decrease in LDL
levels.
[0019] Nicotinic acid, or niacin, also known as vitamin B3, is
effective in reducing triglyceride levels and raising HDL levels
higher than any other anti-cholesterol drug. Nicotinic acid also
lowers LDL-cholesterol.
[0020] Fibric acid derivatives, or fibrates, are used to lower
triglyceride levels and increase HDL when other drugs ordinarily
used for these purposes, such as niacin, are not effective.
[0021] Probucol lowers LDL-cholesterol levels, however, it also
lowers HDL levels. It is generally used for certain genetic
disorders that cause high cholesterol levels, or in cases where
other cholesterol-lowering drugs are ineffective or cannot be
used.
[0022] PCSK9s lower LDL-cholesterol levels via increasing the
cellular level of LDL receptors that reside in the liver.
[0023] Hypolipidemic drugs have had varying degrees of success in
reducing blood lipid; however, none of the hypolipidemic drugs
successfully treats all types of hyperlipidemia. While some
hypolipidemic drugs have been fairly successful, the medical
community has found little conclusive evidence that hypolipidemic
drugs cause regression of atherosclerosis. In addition, all
hypolipidemic drugs have undesirable side effects. As a result of
the lack of success of dietary control, drug therapy and other
therapies, atherosclerosis remains a major cause of death in many
parts of the world.
[0024] New therapies have been used to reduce the amount of lipid
in patients for whom drug and diet therapies were not sufficiently
effective. For example, extracorporeal procedures like
plasmapheresis and LDL-apheresis have been employed and are shown
to be effective in lowering LDL.
[0025] Plasmapheresis therapy or plasma exchange therapy, involves
replacing a patient's plasma with donor plasma or more usually a
plasma protein fraction. Plasmapheresis is a process whereby the
blood plasma is removed from blood cells by a cell separator. The
separator works either by spinning the blood at high speed to
separate the cells from the fluid or by passing the blood through a
membrane with pores so small that only the fluid component of the
blood can pass through. The cells are returned to the person
undergoing treatment, while the plasma is discarded and replaced
with other fluids.
[0026] This treatment has resulted in complications due to the
introduction of foreign proteins and transmission of infectious
diseases. Further, plasmapheresis has the disadvantage of
non-selective removal of all serum lipoproteins, such as VLDL, LDL,
and HDL. Moreover, plasmapheresis can result in several side
effects including allergic reactions in the form of fever, chills,
and rash and possibly even anaphylaxis.
[0027] As described above, it is not desirable to remove HDL, which
is secreted from both the liver and the intestine as nascent,
disk-shaped particles that contain cholesterol and phospholipids.
HDL is believed to play a role in reverse cholesterol transport,
which is the process by which excess cholesterol is removed from
tissues and transported to the liver for reuse or disposal in the
bile.
[0028] In contrast to plasmapheresis, the LDL-apheresis procedure
selectively removes ApoB containing cholesterol, such as LDL, while
retaining HDL. Several methods for LDL-apheresis have been
developed. These techniques include absorption of LDL in
heparin-agarose beads, the use of immobilized LDL-antibodies,
cascade filtration absorption to immobilize dextran sulfate, and
LDL precipitation at low pH in the presence of heparin. Each method
described above is effective in removing LDL. This treatment
process has disadvantages, however, including the failure to
positively affect HDL or to cause a metabolic shift that can
enhance atherosclerosis and other cardiovascular diseases. LDL
apheresis, as its name suggests, merely treats LDL in patients with
severe hyperlipidemia.
[0029] Yet another method of achieving a reduction in plasma
cholesterol in homozygous familial hypercholesterolemia,
heterozygous familial hypercholesterolemia and patients with
acquired hyperlipidemia is an extracorporeal lipid elimination
process, referred to as cholesterol apheresis. In cholesterol
apheresis, blood is withdrawn from a patient, the plasma is
separated from the blood, and the plasma is mixed with a solvent
mixture. The solvent mixture extracts lipids from the plasma.
Thereafter, the delipidated plasma is recombined with the patient's
blood cells and returned to the patient. Using this procedure,
however, results in a modification of the LDL particles, such that
the modified LDL particles could result in increased intensity of
the heart disease. At the same time, this process also resulted in
further delipidation of the HDL particles.
[0030] U.S. Pat. Nos. 7,361,739; 7,375,191; 7,393,826; 8,030,281;
8,048,015; 8,268,787; and 8,637,460, assigned to the Applicant of
the present specification, and herein incorporated by reference in
their entirety, all describe "systems, apparatus and methods for
creating derivatives of at least one form of HDL without
substantially affecting LDL. These derivatives of HDL are particles
with reduced lipid content, particularly reduced cholesterol
content. These particles have the capacity to bind cholesterol and
are administered to a patient to enhance cellular cholesterol
efflux and reduce cholesterol levels in cells, tissues, organs, and
blood vessels".
[0031] U.S. Pat. No. 7,375,191, assigned to the Applicant of the
present specification, and herein incorporated by reference in its
entirety, describes "[a] composition comprising substantially
unmodified low density lipoprotein particles and a particle
derivative of high density lipoprotein particles comprising lipids,
apolipoprotein A-1 and at least one of apolipoprotein
apolipoprotein D or apolipoprotein E, wherein the lipids include
phospholipids, wherein the composition is formed by an
extracorporeal process comprising exposing a biological fluid
comprising low density lipoprotein particles and high density
lipoprotein particles to a lipid removing agent, wherein the
substantially unmodified low density lipoprotein particles are
substantially unmodified as compared to the low density lipoprotein
particles in the biological fluid prior to exposure of the
biological fluid to the lipid removing agent, and wherein the
particle derivative of the high density lipoprotein particles has a
lower content of at least one of the phospholipids or cholesterol
than the high density lipoprotein particles in the biological fluid
prior to exposure of the biological fluid to the lipid removing
agent."
[0032] U.S. Pat. No. 7,361,739, assigned to the Applicant of the
present specification, and herein incorporated by reference in its
entirety, describes "[a] composition comprising a particle
derivative of an HDL particle and a substantially unaffected LDL
particle, the particle derivative comprising a lipid bilayer
comprising phospholipids and a protein shell comprising
apolipoprotein A-1 and apolipoprotein A-2, and at least one of
apolipoprotein apolipoprotein D or apolipoprotein E, wherein the
particle derivative has a lower content of at least one of
phospholipids or cholesterol than the HDL particle, and wherein a
content of at least one of phospholipids or cholesterol in the
substantially unaffected LDL particle is substantially similar to a
content of at least one of phospholipids or cholesterol,
respectively, in an LDL particle; and wherein the composition is
obtained extracorporeally."
[0033] U.S. Pat. No. 7,393,826, assigned to the Applicant of the
present specification, and herein incorporated by reference in its
entirety, describes "[a] selectively delipidated biological fluid
comprising a particle derivative of an HDL particle and a
substantially unmodified LDL particle as compared to an LDL
particle, wherein the selectively delipidated biological fluid is
formed by an extracorporeal selective delipidation process
comprising the step of exposing a biological fluid comprising the
HDL particle and the LDL particle to a lipid removing agent,
wherein the particle derivative of the HDL particle comprises a
lipid bilayer comprising phospholipids and a protein shell
comprising apolipoprotein A-1, apolipoprotein A-2, and at least one
of apolipoprotein apolipoprotein D or apolipoprotein E, and wherein
a cholesterol content of the HDL particle derivative is lower than
a cholesterol content of the HDL particle."
[0034] U.S. Pat. No. 8,030,281, assigned to the Applicant of the
present specification, and herein incorporated by reference in its
entirety, describes "[a] method for making a particle derivative of
at least one form of high density lipoprotein wherein the particle
derivative comprises a protein shell and lipid comprising the steps
of: a. connecting a patient to a device for withdrawing blood; b.
withdrawing blood containing blood cells from the patient; c.
separating the blood cells from the blood to yield a blood fraction
containing high density lipoprotein and low density lipoprotein; d.
separating the low density lipoprotein from the blood fraction; e.
mixing the blood fraction with a solvent which removes lipid
associated with the high density lipoprotein to yield a mixture of
lipid, the solvent, and the particle derivative; and, f separating
the particle derivative, wherein the particle derivative comprises
apolipoprotein A1 and phospholipid, from the lipid and the
solvent."
[0035] U.S. Pat. No. 8,048,015, assigned to the Applicant of the
present specification, and herein incorporated by reference in its
entirety, describes "[a] method of modifying a protein distribution
in a fluid, wherein the protein distribution has a first state, the
first state having alpha high density lipoproteins and pre-beta
high density lipoproteins, comprising the steps of: exposing the
fluid to a lipid removing agent wherein the exposure modifies the
protein distribution from the first state into a second state, the
second state having an increased concentration of pre-beta high
density lipoproteins relative to the first state; and, removing the
lipid removing agent from the fluid, wherein the lipid removing
agent comprises sevoflurane."
[0036] U.S. Pat. No. 8,268,787, assigned to the Applicant of the
present specification, and herein incorporated by reference in its
entirety, describes "[a] method for enhancing cellular cholesterol
efflux in a patient, comprising administering to the patient a
composition comprising a particle derivative of at least one form
of high density lipoprotein, wherein the particle derivative
comprises a protein shell and lipid and is obtained by a process
comprising the steps of: a. connecting a patient to a device for
withdrawing blood; b. withdrawing blood containing blood cells from
the patient; c. separating the blood cells from the blood to yield
a blood fraction containing high density lipoprotein and low
density lipoprotein; d. separating the low density lipoprotein from
the blood fraction; e. mixing the blood fraction with a solvent
which removes lipid associated with the high density lipoprotein to
yield a mixture of lipid, the solvent, and the particle derivative;
and, f. separating the particle derivative from the lipid and the
solvent, wherein the particle derivative comprises apolipoprotein
A1 and phospholipid, and wherein the particle derivative has a
reduced lipid content as compared to the high density lipoprotein
particle that does not have the solvent treatment."
[0037] U.S. Pat. No. 8,637,460, assigned to the Applicant of the
present specification, and herein incorporated by reference in its
entirety, describes "[a] method of modifying a protein distribution
in a fluid, wherein the protein distribution has a first state, the
first state having alpha high density lipoproteins and pre-beta
high density lipoproteins, comprising the steps of: exposing the
fluid to a lipid removing agent wherein the exposure modifies the
protein distribution from the first state into a second state, the
second state having an increased concentration of pre-beta high
density lipoproteins relative to the first state; and, removing the
lipid removing agent from the fluid, wherein the lipid removing
agent comprises a combination of sevoflurane with at least one of
n-butanol, hexanol, ethanol, isoflurane, diisopropyl ether or
trifluoroethane."
[0038] Further, U.S. patent application Ser. Nos. 16/003,926 and
15/876,808 assigned to the Applicant of the present specification,
are also herein incorporated by reference in their entirety.
[0039] Vigorous multi-stage solvent exposure and extraction can
have several drawbacks. It may be difficult to remove a sufficient
amount of solvents from the delipidated plasma in order for the
delipidated plasma to be safely returned to a patient. What is also
needed is a system and a method to process the plasma and solvent
mixture and consequently derive delipidated plasma that can be
provided to a patient, in chronic diseases.
[0040] What are also needed are systems and methods that provide a
simple and improved priming and waste management process. More
specifically, what is needed is a system and method that is capable
of processing both solvent waste and prime waste separately so that
the waste streams can be treated and disposed of appropriately.
SUMMARY
[0041] The following embodiments and aspects thereof are described
and illustrated in conjunction with systems, tools and methods,
which are meant to be exemplary and illustrative, not limiting in
scope.
[0042] In some embodiments, the present specification discloses a
plasma processing system comprising: a source of saline; a first
fluid flow line; a first fluid flow line valve positioned between
the source of saline and the first fluid flow line; a second fluid
flow line; a second fluid flow line valve positioned between the
first fluid flow line and a second fluid flow line; a third fluid
flow line; a pump positioned between the second fluid flow line and
the third fluid flow line; a prime waste container in fluid
communication with the third fluid flow line; a fourth fluid flow
line; a fourth fluid flow line valve positioned between the first
fluid flow line and the fourth fluid flow line; a third fluid flow
path valve positioned along the third fluid flow path; a connector
tube; a solvent extraction device, wherein the connector tube and
solvent extraction device are configured to be alternatively
inserted in a same position along the second fluid flow line; and a
controller, wherein the controller is configured to execute a
plurality of programmatic instructions to open and close each of
the first fluid flow line valve, second fluid flow line valve,
third fluid flow line valve, and fourth fluid flow line valve in a
predetermined sequence.
[0043] Optionally, the solvent extraction device is a charcoal
column.
[0044] Optionally, the first fluid flow path is not in fluid
communication with a source of plasma or a source of solvent.
[0045] Optionally, the plasma processing system further comprises a
separator, wherein the fourth fluid flow line is in fluid
communication with the separator.
[0046] Optionally, the plasma processing system further comprises
an output plasma container, wherein the output plasma container is
in fluid communication with the pump.
[0047] Optionally, the controller is configured to execute a
plurality of programmatic instructions to open and close each of
the first fluid flow line valve, second fluid flow line valve,
third fluid flow line valve, and fourth fluid flow line valve such
that the first fluid flow line, second fluid flow line, and fourth
fluid flow line are filled with saline before the fourth fluid flow
line is closed.
[0048] Optionally, the controller is configured to execute a
plurality of programmatic instructions to open and close each of
the first fluid flow line valve, second fluid flow line valve,
third fluid flow line valve, and fourth fluid flow line valve such
that the first fluid flow line, second fluid flow line, and fourth
fluid flow line are filled with saline before the third fluid flow
line is opened.
[0049] Optionally, the controller is configured to execute a
plurality of programmatic instructions to activate the pump and
open and close each of the first fluid flow line valve, second
fluid flow line valve, third fluid flow line valve, and fourth
fluid flow line valve such that fluid in the first fluid flow line
and second fluid flow line is directed to the prime waste
container.
[0050] In some embodiments, the present specification is directed
towards a plasma processing system comprising: a source of saline;
a plurality of fluid flow lines, wherein the plurality of fluid
flow lines comprise a first fluid flow line, a second fluid flow
line, a third fluid flow line, and a fourth fluid flow line; a
plurality of fluid flow line valves positioned along each of the
plurality of fluid flow lines; a connector tube; a solvent
extraction device, wherein the connector tube and solvent
extraction device are configured to be alternatively inserted in a
same position along the second fluid flow line; and a controller,
wherein the controller is configured to execute a plurality of
programmatic instructions to open and close each of the plurality
of fluid flow line valves in a predetermined sequence.
[0051] Optionally, the solvent extraction device is a charcoal
column.
[0052] Optionally, the plasma processing system further comprises a
separator, wherein the fourth fluid flow line is in fluid
communication with the separator.
[0053] Optionally, the plurality of fluid flow line valves comprise
a first fluid flow line valve positioned between the source of
saline and the first fluid flow line, a second fluid flow line
valve positioned between the first fluid flow line and the second
fluid flow line, a fourth fluid flow line valve positioned between
the first fluid flow line and the fourth fluid flow line, and a
third fluid flow path valve positioned along the third fluid flow
path.
[0054] Optionally, the plasma processing system further comprises a
pump positioned between the second fluid flow line and the third
fluid flow line.
[0055] Optionally, the plasma processing system further comprises a
prime waste container in fluid communication with the third fluid
flow line.
[0056] Optionally, the controller is configured to execute a
plurality of programmatic instructions to open and close each of
the first fluid flow line valve, second fluid flow line valve,
third fluid flow line valve, and fourth fluid flow line valve such
that the first fluid flow line, second fluid flow line, and fourth
fluid flow line are filled with saline before the fourth fluid flow
line is closed.
[0057] Optionally, the controller is configured to execute a
plurality of programmatic instructions to open and close each of
the first fluid flow line valve, second fluid flow line valve,
third fluid flow line valve, and fourth fluid flow line valve such
that the first fluid flow line, second fluid flow line, and fourth
fluid flow line are filled with saline before the third fluid flow
line is opened.
[0058] Optionally, the controller is configured to execute a
plurality of programmatic instructions to activate the pump and
open and close each of the first fluid flow line valve, second
fluid flow line valve, third fluid flow line valve, and fourth
fluid flow line valve such that fluid in the first fluid flow line
and second fluid flow line is directed to the prime waste
container.
[0059] In some embodiments, the present specification discloses a
device for priming a plasma processing system comprising: a source
of a first fluid; a first valve; a second valve; a first pump; a
prime waste container; a third valve; a fourth valve; a fifth
valve; a source of a second fluid; a first fluid flow path, wherein
the first fluid flow path is defined by the source of a first
fluid, the first valve positioned between the source of the first
fluid and a first fluid flow line, the second valve positioned
between the first fluid flow line and a second fluid flow line, the
first pump positioned between the second fluid flow line and a
third fluid flow line, and the prime waste container in fluid
communication with the third fluid flow line; and a controller,
wherein the controller is configured to execute a plurality of
programmatic instructions to: open the first valve, thereby
allowing a flow of fluid from the source of the first fluid to the
prime waste container, along the first fluid flow path; close the
second valve, thereby preventing a flow of fluid from the source of
the first fluid to the second fluid flow line, third fluid flow
line, and the prime waste container; close the first valve, thereby
preventing a flow of the first fluid to the first fluid flow line
from the source of the first fluid; open the third valve, wherein
the third valve is positioned between the first fluid flow line and
a fourth fluid flow line; open the fourth valve, wherein the fourth
valve is positioned between the source of the second fluid and the
first fluid flow line, thereby enabling a flow of fluid along a
second fluid flow path; and close the third valve and opening the
second valve and a fifth valve, wherein the fifth valve is
positioned along a third fluid flow path, thereby enabling a flow
of fluid along the third fluid flow path to the prime waste
container.
[0060] Optionally, the controller is further configured to execute
a plurality of programmatic instructions to close the first valve,
thereby preventing a flow of the first fluid to the first fluid
flow line from the source of the first fluid. Optionally, the first
fluid is saline. Optionally, the second fluid is saline.
[0061] Optionally, the device further comprises a connector tube
positioned along the second fluid flow line.
[0062] Optionally, the device further comprises a solvent
extraction device positioned along the second fluid flow line.
Optionally, the solvent extraction device is a charcoal column.
[0063] Optionally, the device further comprises a connector tube
positioned along the second fluid flow line and a solvent
extraction device configured to be inserted in a same position as
the connector tube when the connector tube is removed from the
second fluid flow line.
[0064] Optionally, the solvent extraction device is a charcoal
column.
[0065] Optionally, the first fluid flow path is not in fluid
communication with a source of plasma or a source of solvent.
[0066] Optionally, the device further comprises a separator,
wherein the fourth fluid flow line is in fluid communication with
the separator.
[0067] In some embodiments, the present specification discloses a
device for priming a plasma processing system comprising: a source
of saline; a first fluid flow line; a first fluid flow line valve
positioned between the source of saline and the first fluid flow
line; a second fluid flow line; a second fluid flow line valve
positioned between the first fluid flow line and a second fluid
flow line; a third fluid flow line; a pump positioned between the
second fluid flow line and the third fluid flow line; a prime waste
container in fluid communication with the third fluid flow line; a
fourth fluid flow line; a fourth fluid flow line valve positioned
between the first fluid flow line and the fourth fluid flow line; a
third fluid flow path valve positioned along the third fluid flow
path; and a controller, wherein the controller is configured to
execute a plurality of programmatic instructions to: close the
second fluid flow line valve, thereby preventing a flow of fluid
from the source of saline to the second fluid flow line, third
fluid flow line, and the prime waste container; open the fourth
fluid flow line valve; open the first fluid flow line valve,
thereby enabling a flow of saline along the first fluid flow line
and the second fluid flow path; and close the fourth fluid flow
line valve and open the second fluid flow line valve and the third
fluid flow path valve, thereby enabling a flow of fluid along the
third fluid flow path to the prime waste container.
[0068] Optionally, the device further comprises a connector tube
positioned along the second fluid flow line.
[0069] Optionally, the device further comprises a solvent
extraction device positioned along the second fluid flow line.
Optionally, the solvent extraction device is a charcoal column.
[0070] Optionally, the device further comprises a connector tube
positioned along the second fluid flow line and a solvent
extraction device configured to be inserted in a same position as
the connector tube when the connector tube is removed from the
second fluid flow line. Optionally, the solvent extraction device
is a charcoal column.
[0071] Optionally, the first fluid flow path is not in fluid
communication with a source of plasma or a source of solvent.
[0072] Optionally, the device further comprises a separator,
wherein the fourth fluid flow line is in fluid communication with
the separator.
[0073] Optionally, the device further comprises an output plasma
container, wherein the output plasma container is in fluid
communication with the pump.
[0074] In some embodiments, the present specification discloses a
method of priming a plasma processing system comprising at least a
first fluid flow path, a second fluid flow path, third fluid flow
path, and a fourth fluid flow path, comprising: flushing a first
fluid circuit, wherein the first fluid circuit is defined by a
source of a first fluid, a first valve positioned between the
source of the first fluid and the first fluid flow path, a second
valve positioned between the first fluid flow path and the second
fluid flow path, a first pump positioned between the second fluid
flow path and the third fluid flow path, and a first waste
container in fluid communication with the third fluid flow path;
closing the second valve, thereby preventing a flow of fluid to the
second fluid flow path, third fluid flow path, and first waste
container; closing the first valve, thereby preventing a flow of
the first fluid to the first fluid flow path from the source of the
first fluid; opening a third valve, wherein the third valve is
positioned between the first fluid flow path and the fourth fluid
flow path; opening a fourth valve, wherein the fourth valve is
positioned between a source of a second fluid and the first fluid
flow path; and opening the second valve, thereby enabling a flow of
fluid to the second fluid flow path, third fluid flow path, and
first waste container.
[0075] Optionally, the first fluid is saline.
[0076] Optionally, the second fluid is saline.
[0077] Optionally, the first fluid circuit is not in fluid
communication with a source of plasma, a source of solvent, or an
output plasma container.
[0078] Optionally, the plasma processing system further comprises a
connector tube positioned along the second fluid flow path.
Optionally, the method further comprises, after opening the second
valve, clamping the second fluid flow path and removing the
connector tube. Optionally, the method further comprises, after
removing the connector tube, inserting a solvent extraction device
in place of the removed connector tube. Optionally, the solvent
extraction device is a charcoal column.
[0079] Optionally, plasma processing system further comprises a
fifth valve positioned between the third fluid flow path and the
first waste container.
[0080] Optionally, the fourth fluid flow path is in fluid
communication with a separator.
[0081] The present specification also discloses a method of priming
a plasma processing system comprising at least a first fluid flow
path, a second fluid flow path, third fluid flow path, and a fourth
fluid flow path, comprising: flushing a first fluid circuit,
wherein the first fluid circuit is defined by a source of a first
fluid, a first valve positioned between the source of the first
fluid and the first fluid flow path, a second valve positioned
between the first fluid flow path and the second fluid flow path, a
first pump positioned between the second fluid flow path and the
third fluid flow path, and a first waste container in fluid
communication with the third fluid flow path; and flushing a second
fluid circuit, wherein the second fluid circuit is defined by a
source of a second fluid, a third valve, wherein the third valve is
positioned between the first fluid flow path and the fourth fluid
flow path, and a fourth valve, wherein the fourth valve is
positioned between a source of a second fluid and the first fluid
flow path, by closing the second valve, thereby preventing a flow
of fluid to the second fluid flow path, third fluid flow path, and
first waste container, closing the first valve, thereby preventing
a flow of the first fluid to the first fluid flow path from the
source of the first fluid, opening the third valve, and opening the
fourth valve.
[0082] Optionally, the first fluid is saline and the second fluid
is saline.
[0083] Optionally, the first fluid circuit is not in fluid
communication with a source of plasma, a source of solvent, or an
output plasma container.
[0084] Optionally, the plasma processing system further comprises a
connector tube positioned along the second fluid flow path.
[0085] Optionally, the method further comprises, after closing the
second valve, waiting a period of time and then opening the second
valve, thereby enabling a flow of fluid to the second fluid flow
path, third fluid flow path, and first waste container.
[0086] Optionally, the method further comprises, after opening the
second valve, clamping the second fluid flow path and removing the
connector tube. Optionally, the method further comprises, after
removing the connector tube, inserting a solvent extraction device
in place of the removed connector tube. Optionally, the solvent
extraction device is a charcoal column.
[0087] Optionally, the plasma processing system further comprises a
fifth valve positioned between the third fluid flow path and the
first waste container.
[0088] Optionally, the fourth fluid flow path is in fluid
communication with a separator.
[0089] The present specification also discloses a method for
treating plasma using an apparatus to treat the plasma with a
solvent, the method comprising: configuring the apparatus to
separate a solvent waste and a prime waste; priming the apparatus
with a priming fluid, the priming resulting in priming waste,
wherein the priming waste is collected in a container configured to
collect prime waste; installing a solvent extraction device within
the apparatus; priming the apparatus with the solvent extraction
device, the priming resulting in priming waste, wherein the priming
waste is collected in the container configured to collect prime
waste; introducing the plasma and the solvent in to a mixing
device; mixing the plasma and the solvent; separating the plasma
and the solvent, wherein the solvent is removed from the plasma
into a container configured to collect solvent waste; and
extracting remaining solvent from the plasma by transporting the
separated plasma through the solvent extraction device.
[0090] Optionally, the solvent is at least one or more of a
combination of n-butanol, ethyl acetate, dichloromethane,
chloroform, isoflurane, sevoflurane (1,1, 1,3,
3,3-hexafluoro-2-(fluoromethoxy) propane-d3),
perfluorocyclohexanes, trifluoroethane, and cyclofluorohexanol.
[0091] Optionally, the solvent is a lipid removing agent which
removes lipids to yield a mixture of lipid, the lipid removing
agent, modified high density lipoprotein, and the low density
lipoprotein, wherein the modified high density lipoprotein is a
delipidated high density lipoprotein.
[0092] Optionally, separating the plasma and the solvent comprises
separating the modified high density lipoprotein and the low
density lipoprotein from the lipid and the lipid removing
agent.
[0093] Optionally, the step of separating the plasma and the
solvent comprises using gravity.
[0094] Optionally, configuring the apparatus to separate a solvent
waste and a prime waste comprises configuring a first waste
container to collect solvent waste and a second waste container,
separate from the first waste container, to collect prime
waste.
[0095] Optionally, priming the apparatus with a priming fluid
further comprises attaching a prime connector tube in the
apparatus, wherein the prime connector tube is replaced by the
solvent extraction device.
[0096] The present specification also discloses a method of using
an apparatus to modify protein distribution in a fluid, wherein the
method comprises: priming the apparatus with a priming fluid, the
priming resulting in priming waste, wherein the priming waste is
collected in a second waste container configured to collect prime
waste; installing a solvent extraction device within the apparatus;
priming the apparatus with the solvent extraction device, the
priming resulting in priming waste, wherein the priming waste is
collected in the second container; inputting a plasma in to a first
fluid container; opening a first valve to direct flow from the
first fluid container to a mixing device; inputting a solvent in to
a second fluid container; opening a second valve to direct flow
from the second fluid container to the mixing device; mixing the
plasma and the solvent in the mixer for a first predetermined
period of time; after the first predetermined period of time,
opening a third valve to direct the plasma and the solvent mixture
to a funnel-shaped bag separator; separating the plasma and the
solvent in the separator for a second predetermined period of time;
after the second predetermined period of time, opening a fourth
valve to direct flow of separated solvent from the separator in to
a first waste container configured to collect solvent waste;
opening a fifth valve to direct flow of separated plasma from the
separator in to the solvent extraction device; closing a sixth
valve to inhibit flow of separated plasma from the solvent
extraction device in to the second waste container; and opening a
seventh valve to direct flow of separated plasma from the solvent
extraction device in to a third fluid container configured to
collect separated plasma.
[0097] Optionally, the opening the third valve to direct the plasma
and the solvent mixture to the funnel-shaped bag separator results
in gravity-directed flow of the plasma and the solvent mixture.
[0098] Optionally, the method further comprises pumping the priming
fluid to direct flow of the priming fluid towards the second
container, and pumping the separated plasma to direct flow of the
separated plasma in to the solvent extraction device and the third
fluid container.
[0099] Optionally, opening the fourth valve to direct flow of
separated solvent from the separator comprises directing the flow
through a cone-shaped bottom of the separator.
[0100] Optionally, the solvent extraction device is a charcoal
column.
[0101] The present specification also discloses a method for mixing
a plasma and a solvent in a mixing device, to modify protein
distribution in the plasma, the method comprising: introducing a
first volume of the plasma in to the mixing device, wherein said
mixing device comprises at least one of a mixing bag, a mixer and a
platform positioned above the mixer for said mixing bag to be
placed upon; introducing a second volume of the solvent in to the
mixing device; and mixing the first volume of the plasma and the
second volume of the solvent in the mixing device, wherein the
mixing device has a set of features; and varying at least one of
the first volume, the second volume, the plasma, the solvent, the
mixing device, and the set of features of the mixing device, to
vary the extent of modification of protein distribution in the
plasma.
[0102] Optionally, varying the mixing device comprises using a
mixer that is one of an orbital mixer, a vortex mixer, a rotating
table mixer, and a coiled tube mixer.
[0103] Optionally, varying the mixing device comprises varying an
amount of energy applied to the mixer.
[0104] Optionally, varying the mixing device comprises varying a
shape of the mixing bag.
[0105] Optionally, varying the mixing device comprises varying an
angle at which the platform is positioned.
[0106] Optionally, varying the mixing device comprises varying a
speed of operation of the mixer.
[0107] Optionally, a duration of the mixing is varied.
[0108] Optionally, the varying of the first volume and of the
second volume comprises varying a ratio of the first volume to the
second volume.
[0109] Optionally, varying the plasma comprises selecting one of a
human plasma, a bovine plasma, a normal plasma, and a lipemic IV
plasma.
[0110] Optionally, a ration of the constituents of the solvent is
varied.
[0111] Optionally, a ration of the plasma to solvent is varied.
[0112] The aforementioned and other embodiments of the present
specification shall be described in greater depth in the drawings
and detailed description provided below.
BRIEF DESCRIPTION OF THE DRAWINGS
[0113] These and other features and advantages of the present
invention will be appreciated, as they become better understood by
reference to the following detailed description when considered in
connection with the accompanying drawings, wherein:
[0114] FIG. 1 is a schematic representation of a prior art system
comprising a plurality of components used in accordance with some
embodiments of the present specification to achieve the processes
disclosed herein;
[0115] FIG. 2 is a flow chart illustrating an exemplary process for
treating cardiovascular diseases using the system of FIG. 1, in
accordance with some embodiments of the present specification;
[0116] FIG. 3A is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0117] FIG. 3B is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0118] FIG. 3C is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0119] FIG. 3D is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0120] FIG. 3E is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0121] FIG. 3F is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0122] FIG. 3G is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0123] FIG. 3H is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0124] FIG. 3I is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0125] FIG. 3J is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0126] FIG. 3K is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0127] FIG. 3L is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0128] FIG. 3M is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0129] FIG. 3N is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0130] FIG. 3O is a schematic representation of the system
illustrating the implementation of the process described in FIG. 2,
in accordance with some embodiments of the present
specification;
[0131] FIG. 4 is a table showing the effect on the reduction of
lipids resulting from variation in different chemical and
mechanical parameters involved in implementing various embodiments
in accordance with the present specification;
[0132] FIG. 5 is a table listing another exemplary set of variables
that affect the delipidation process and outcome;
[0133] FIG. 6 is a table providing another exemplary set of
variables that may be used for normal plasma and lipemic IV plasma
using different solvents and different methods of separation;
[0134] FIG. 7 illustrates an exemplary mixing device, in accordance
with embodiments described in context of FIG. 3I;
[0135] FIG. 8A illustrates a side view of shaker angle brackets
that are used to a position mixing device within a system, in
accordance with some embodiments of the present specification;
[0136] FIG. 8B illustrates another side view of shaker angle
brackets that are used to position a mixing device within a system,
in accordance with some embodiments of the present specification;
and
[0137] FIG. 8C illustrates a perspective view of shaker angle
brackets that are used to position a mixing device within a system,
in accordance with some embodiments of the present
specification.
DETAILED DESCRIPTION
[0138] In some embodiments, the present specification is directed
towards systems, apparatuses and methods for removing lipid from
.alpha.-High Density Lipoprotein (.alpha.-HDL) particles derived
primarily from plasma of the patient thereby creating modified HDL
particles (also referred to as delipidated HDL) with reduced lipid
content, particularly reduced cholesterol content. Embodiments of
the present specification create these modified HDL particles with
reduced lipid content without substantially modifying LDL
particles. Embodiments of the present specification modify original
.alpha.-HDL particles (present in delipidated plasma) to yield
modified HDL particles that have an increased concentration of
pre-.beta. HDL relative to the original HDL. The modified HDL, with
a concentrated solution of pre-.beta. HDL is administered to the
patient to enhance cellular cholesterol efflux and treat
cardiovascular diseases and/or other lipid-associated diseases.
[0139] The treatment processes of the present specification renders
the methods and systems of the present specification more effective
in treating cardiovascular diseases including Homozygous Familial
Hypercholesterolemia (HoFH), Heterozygous Familial
Hypercholesterolemia (HeFH), Ischemic stroke, Coronary Artery
Disease (CAD), Acute Coronary Syndrome (ACS), peripheral arterial
disease (PAD), Renal Arterial Stenosis (RAS), and for treating the
progression of Alzheimer's Disease.
[0140] Embodiments of the present specification provide systems and
methods to achieve the above objectives. Systems and methods are
provided where plasma and solvent(s) are introduced into a
specially designed mixing bag in precise quantities and volumetric
ratios. The solvent and plasma are then mixed in an orbital fashion
for a prescribed period, resulting in delipidation. The mixture is
then drained into a separator bag. Each batch is mixed and drained
into the separator bag until the input plasma is fully processed.
When the separator bag reaches capacity, excess solvent is drained
to a solvent waste bag.
[0141] The timed suspension in the separator bag separates the
plasma and solvent into distinct fractions so the solvent can be
drained into the solvent waste bag. Some solvent, however, remains
dissolved in the plasma. This residual solvent is substantially
removed by passing the plasma through a specially-designed charcoal
column. The output plasma contains selectively delipidated HDL with
substantially unchanged or undelipidated LDL.
[0142] The present specification is directed towards multiple
embodiments. The following disclosure is provided in order to
enable a person having ordinary skill in the art to practice the
invention. Language used in this specification should not be
interpreted as a general disavowal of any one specific embodiment
or used to limit the claims beyond the meaning of the terms used
therein. The general principles defined herein may be applied to
other embodiments and applications without departing from the
spirit and scope of the invention. Also, the terminology and
phraseology used is for the purpose of describing exemplary
embodiments and should not be considered limiting. Thus, the
present invention is to be accorded the widest scope encompassing
numerous alternatives, modifications and equivalents consistent
with the principles and features disclosed. For purpose of clarity,
details relating to technical material that is known in the
technical fields related to the invention have not been described
in detail so as not to unnecessarily obscure the present invention.
In the description and claims of the application, each of the words
"comprise" "include" and "have", and forms thereof, are not
necessarily limited to members in a list with which the words may
be associated.
[0143] It should be noted herein that any feature or component
described in association with a specific embodiment may be used and
implemented with any other embodiment unless clearly indicated
otherwise.
[0144] The term "fluid" may be defined as fluids from animals or
humans that contain lipids or lipid containing particles, fluids
from culturing tissues and cells that contain lipids and fluids
mixed with lipid-containing cells. For purposes of this invention,
decreasing the amount of lipids in fluids includes decreasing
lipids in plasma and particles contained in plasma, including but
not limited to HDL particles. Fluids include, but are not limited
to: biological fluids; such as blood, plasma, serum, lymphatic
fluid, cerebrospinal fluid, peritoneal fluid, pleural fluid,
pericardial fluid, various fluids of the reproductive system
including, but not limited to, semen, ejaculatory fluids,
follicular fluid and amniotic fluid; cell culture reagents such as
normal sera, fetal calf serum or serum derived from any animal or
human; and immunological reagents, such as various preparations of
antibodies and cytokines from culturing tissues and cells, fluids
mixed with lipid-containing cells, and fluids containing
lipid-containing organisms, such as a saline solution containing
lipid-containing organisms. A preferred fluid treated with the
methods of the present invention is plasma. Arrows on the tubing
segments in the figures represent fluid flow or the movement of
fluid while the absence of arrows represents no fluid flow or
movement. Patterns within the tubing segments in the figures
represent fluid within the tubing while the absence of patterns
represents no fluid within that segment of tubing.
[0145] The term "lipid" may be defined as any one or more of a
group of fats or fat-like substances occurring in humans or
animals. The fats or fat-like substances are characterized by their
insolubility in water and solubility in organic solvents. The term
"lipid" is known to those of ordinary skill in the art and
includes, but is not limited to, complex lipid, simple lipid,
triglycerides, fatty acids, glycerophospholipids (phospholipids),
true fats such as esters of fatty acids, glycerol, cerebrosides,
waxes, and sterols such as cholesterol and ergosterol.
[0146] The term "extraction solvent" may be defined as one or more
solvents used for extracting lipids from a fluid or from particles
within the fluid. This solvent enters the fluid and remains in the
fluid until removed by other subsystems. Suitable extraction
solvents include solvents that extract or dissolve lipid, including
but not limited to phenols, hydrocarbons, amines, ethers, esters,
alcohols, halohydrocarbons, halocarbons, and combinations thereof.
Examples of suitable extraction solvents are ethers, esters,
alcohols, halohydrocarbons, or halocarbons which include, but are
not limited to di-isopropyl ether (DIPE), which is also referred to
as isopropyl ether, diethyl ether (DEE), which is also referred to
as ethyl ether, lower order alcohols such as butanol, especially
n-butanol, ethyl acetate, dichloromethane, chloroform, isoflurane,
sevoflurane (1,1, 1,3, 3,3-hexafluoro-2-(fluoromethoxy)
propane-d3), perfluorocyclohexanes, trifluoroethane,
cyclofluorohexanol, and combinations thereof.
[0147] The term "patient" refers to animals and humans, which may
be either a fluid source to be treated with the methods of the
present invention or a recipient of derivatives of HDL particles
and or plasma with reduced lipid content.
[0148] The term "HDL particles" encompasses several types of
particles defined based on a variety of methods such as those that
measure charge, density, size and immuno-affinity, including but
not limited to electrophoretic mobility, ultracentrifugation,
immunoreactivity and other methods known to one of ordinary skill
in the art. Such HDL particles include but are not limited to the
following: .alpha.-HDL, pre-.beta. HDL (including pre-.beta.1 HDL,
pre-.beta.2 HDL and pre-.beta.3HDL), HDL2 (including HDL2a and
HDL2b), HDL3, VHDL, LpA-I, LpA-II, LpA-I/LpA-II (for a review see
Barrans et al., Biochemica Biophysica Acta 1300; 73-85, 1996).
Accordingly, practice of the methods of the present invention
creates modified HDL particles. These modified derivatives of HDL
particles may be modified in numerous ways including but not
limited to changes in one or more of the following metabolic and/or
physico-chemical properties (for a review see Barrans et al. ,
Biochemica Biophysica Acta 1300; 73-85, 1996); molecular mass
(kDa); charge; diameter; shape; density; hydration density;
flotation characteristics; content of cholesterol; content of free
cholesterol; content of esterified cholesterol; molar ratio of free
cholesterol to phospholipids; immuno-affinity; content, activity or
helicity of one or more of the following enzymes or proteins:
Apo-AI, Apo-AII, ApoD, ApoE, ApoJ, ApoA-IV, cholesterol ester
transfer protein (CETP), lecithin; cholesterol acyltransferase
(LCAT); capacity and/or rate for cholesterol binding, capacity
and/or rate for cholesterol transport.
[0149] The terms "modified high density lipoprotein" and
"delipidated high density lipoprotein" may be used interchangeably
and refer to reduced lipid blood products, and in particular, high
density lipoproteins having a reduced lipid content, that may be
contained within the resultant plasma once a delipidation process
has been performed. Similarly, the term "treated plasma" refers to
the resultant plasma once a delipidation process has been
performed.
[0150] FIG. 1 illustrates an exemplary prior art system and its
components used to achieve the methods of the present
specification. The figure depicts an exemplary basic component flow
diagram defining elements of the HDL modification system 100.
Embodiments of the components of system 100 are utilized after
obtaining a blood fraction from a patient or another individual
(donor). The plasma, separated from the blood is brought in a
sterile bag to system 100 for further processing. The plasma may be
separated from blood using a known plasmapheresis device. The
plasma may be collected from the patient into a sterile bag using
standard apheresis techniques. The plasma is then brought in the
form of a fluid input to system 100 for further processing. In
embodiments, system 100 is not connected to the patient at any time
and is a discrete, stand-along system for delipidating plasma. The
patient's plasma is processed by system 100 and brought back to the
patient's location to be reinfused back into the patient. In
alternate embodiments, the system may be a continuous flow system
that is connected to the patient in which both plasmapheresis and
delipidation are performed in an excorporeal, parallel system and
the delipidated plasma product is returned to the patient.
[0151] A fluid input 105 (containing blood plasma) is provided and
connected via tubing to a mixing device 120. A solvent input 110 is
provided and also connected via tubing to mixing device 120. In
embodiments, valves 115, 116 are used to control the flow of fluid
from fluid input 105 and solvent from solvent input 110
respectively. It should be appreciated that the fluid input 105
contains any fluid that includes HDL particles, including plasma
having LDL particles or devoid of LDL particles, as discussed
above. It should further be appreciated that solvent input 110 can
include a single solvent, a mixture of solvents, or a plurality of
different solvents that are mixed at the point of solvent input
110. While depicted as a single solvent container, solvent input
110 can comprise a plurality of separate solvent containers.
Embodiments of types of solvents that may be used are discussed
subsequently.
[0152] Mixer 120 mixes fluid from fluid input 105 and solvent from
solvent input 110 to yield a fluid-solvent mixture. In some
embodiments, mixer 120 is capable of using a shaker bag mixing
method with the input fluid and input solvent in a plurality of
batches, such as 1, 2, 3 or more batches. In alternative
embodiments, other known methods of mixing are utilized. Once
formed, the fluid-solvent mixture is directed, through tubing and
controlled by at least one valve 115a, to a separator 125. In an
embodiment, separator 125 is capable of performing bulk solvent
separation through gravity separation in a funnel-shaped bag.
[0153] In separator 125, the fluid-solvent mixture separates into a
first layer and second layer. The first layer comprises a mixture
of solvent and lipid that has been removed from the HDL particles.
Typically, the solvent is heavier than the plasma and therefore the
solvent settles at the bottom of separator 125, and the delipidated
plasma is at the top. In embodiments, the density/specific gravity
of solvent is approximately 1.5 times greater than that of the
plasma fluid. In embodiments, separator 125 is conical or V-shaped.
Once the solvent settles at the bottom, it can be easily drained
from separator 125 while the plasma fluid containing HDL particles
is retained. The first layer is transported through a valve 115b to
a first waste container 135. The second layer comprises a mixture
of residual solvent, modified HDL particles, and other elements of
the input fluid. One of ordinary skill in the art would appreciate
that the composition of the first layer and the second layer would
differ based upon the nature of the input fluid. Once the first and
second layers separate in separator 125, the second layer is
transported through tubing to a solvent extraction device 140. In
an embodiment, a pressure sensor (not shown) and valve 130 is
positioned in the flow stream to control the flow of the second
layer to solvent extraction device 140.
[0154] The opening and closing of valves 115, 116 to enable the
flow of fluid from input containers 105, 110 may be timed using
mass balance calculations derived from weight determinations of the
fluid inputs 105, 110, and separator 125. For example, valve 115b
between separator 125 and first waste container 135 and valve 130
between separator 125 and solvent extraction device 140 open after
the input masses (fluid and solvent) substantially balances with
the mass in separator 125 and a sufficient period of time has
elapsed to permit separation between the first and second layers.
Depending on what solvent is used, and therefore which layer
settles to the bottom of separator 125, either valve 115b between
separator 125 and first waste container 135 is opened or valve 130
between separator 125 and solvent extraction device 140 is opened.
One of ordinary skill in the art would appreciate that the timing
of the opening is dependent upon how much fluid is in the first and
second layers and would further appreciate that it is preferred to
keep valve 115b between separator 125 and first waste container 135
open just long enough to remove all of the first layer and some of
the second layer, thereby ensuring that as much solvent as possible
has been removed from the fluid being sent to solvent extraction
device 140.
[0155] In embodiments, an infusion grade fluid ("IGF") may be
employed via one or more inputs 160 which are in fluid
communication with the fluid path 121 leading from separator 125 to
solvent extraction device 140 for priming. In an embodiment, saline
is employed as the infusion grade priming fluid in at least one of
inputs 160. In an embodiment, 0.9% sodium chloride (saline) is
employed. In other embodiments, glucose may be employed as the
infusion grade priming fluid in any one of inputs 160.
[0156] A plurality of valves 115c and 115d are also incorporated in
the flow stream from glucose input 155 and saline input 160
respectively, to the tubing providing the flow path 121 from
separator 125 to solvent extraction device 140. Infusion grade
fluid such as saline and/or glucose is incorporated into
embodiments of the present specification in order to prime solvent
extraction device 140 prior to operation of the system. In
embodiments, saline is used to prime most of the fluid
communication lines and solvent extraction device 140. If priming
is not required, the infusion grade fluid inputs are not employed.
Where such priming is not required, the glucose and saline inputs
are not required. In an embodiment, priming is not required in the
lines between a second waste container 165 and output container
145. Also, one of ordinary skill in the art would appreciate that
the glucose and saline inputs can be replaced with other primers if
required by the solvent extraction device 140.
[0157] In some embodiments, solvent extraction device 140 is a
charcoal column designed to remove the specific solvent used in
solvent input 110. Exemplary solvent extraction device 140 includes
but is not limited to an Asahi Hemosorber.TM. charcoal column or
the Baxter/Gambro Adsorba.TM. 300C charcoal column or any other
charcoal column that is employed in blood hemoglobin perfusion
procedures. In embodiments, it should be noted that if the charcoal
column is pre-primed with glucose, it will limit the amount of
glucose removed from plasma because the free glucose in the priming
agent will bind to glucose sites in the charcoal column, limiting
its ability to absorb more glucose. A pump 150 is used to move the
second layer from separator 125, through solvent extraction device
140, and to output container 145, through a U-shaped configuration.
In embodiments, pump 150 is a rotary peristaltic pump, such as a
Masterflex Model 77201-62.
[0158] The first layer is directed to waste container 135 that is
in fluid communication with separator 125 through tubing and at
least one valve 115b. Additionally, other waste, if generated, can
be directed from the fluid path connecting solvent extraction
device 140 and output container 145 to second waste container 165.
Optionally, in an embodiment, a valve 115f is included in the path
from the solvent extraction device 140 to the output container 145.
Optionally, in an embodiment, a valve 115g is included in the path
from the solvent extraction device 140 to the second waste
container 165.
[0159] In an embodiment of the present specification, gravity is
used, wherever practical, to move fluid through each of the
plurality of components. For example, gravity is used to drain
input plasma 105 and input solvent 110 into mixer 120. Where mixer
120 comprises a shaker bag and separator 125 comprises a funnel
bag, fluid is moved from the shaker bag to the funnel bag and,
subsequently, to first waste container 135, if appropriate, using
gravity.
[0160] Suitable materials for use in any of the apparatus
components, including bags and tubing, as described herein include
materials that are biocompatible, approved for medical applications
that involve contact with internal body fluids, and in compliance
with U.S. PVI or ISO 10993 standards. Further, the materials do not
substantially degrade from, for instance, exposure to the solvents
used in the present invention, during at least a single use. The
materials are sterilisable by radiation, steam or ethylene oxide
(EtO) sterilization. Such suitable materials are capable of being
formed into objects using conventional processes, such as, but not
limited to, extrusion, injection molding and others. Materials
meeting these requirements include, but are not limited to, nylon,
polypropylene, polycarbonate, acrylic, polysulfone, polyvinylidene
fluoride (PVDF), fluoroelastomers such as VITON, available from
DuPont Dow Elastomers L.L.C., thermoplastic elastomers such as
SANTOPRENE, available from Monsanto, polyurethane, polyvinyl
chloride (PVC), polytetrafluoroethylene (PTFE), polyphenylene ether
(PFE), perfluoroalkoxy copolymer (PFA), which is available as
TEFLON PFA from E.I. du Pont de Nemours and Company, and
combinations thereof.
[0161] Valves 115, 115a, 115b, 115c, 115d, 115e, 115f, 115g, 116
and any other valve used in each embodiment may be composed of, but
are not limited to, pinch, globe, ball, gate or other conventional
valves. In some embodiments, the valves are occlusion valves such
as Acro Associates' Model 955 valve. However, the present
specification is not limited to a valve having a particular style.
Further, the components of each system described in accordance with
embodiments of the present specification may be physically coupled
together or coupled together using conduits that may be composed of
flexible or rigid pipe, tubing or other such devices known to those
of ordinary skill in the art.
[0162] In an additional embodiment, not shown in FIG. 1, the output
fluid in output container 145 is subjected to a solvent detection
system, or lipid removing agent detection system, to determine if
any solvent, or other undesirable component, is in the output
fluid. In embodiments, a solvent sensor is only employed in a
continuous flow system. In one embodiment, the output fluid is
subjected to sensors that are capable of determining the
concentrations of solvents introduced in the solvent input, such as
n-butanol or di-isopropyl ether. In embodiments, the sensors are
capable of providing such concentration information on a real-time
basis and without having to physically transport a sample of the
output fluid, or air in the headspace, to a remote device. The
resultant separated modified HDL particles are then introduced to
the bloodstream of the patient.
[0163] In one embodiment, molecularly imprinted polymer technology
is used to enable surface acoustic wave sensors. A surface acoustic
wave sensor receives an input, through some interaction of its
surface with the surrounding environment, and yields an electrical
response, generated by the piezoelectric properties of the sensor
substrate. To enable the interaction, molecularly imprinted polymer
technology is used. Molecularly imprinted polymers are plastics
programmed to recognize target molecules, like pharmaceuticals,
toxins or environmental pollutants, in complex biological samples.
The molecular imprinting technology is enabled by the
polymerization of one or more functional monomers with an excess of
a crosslinking monomer in presence of a target template molecule
exhibiting a structure similar to the target molecule that is to be
recognized, i.e. the target solvent.
[0164] The use of molecularly imprinted polymer technology to
enable surface acoustic wave sensors can be made more specific to
the concentrations of targeted solvents and are capable of
differentiating such targeted solvents from other possible
interferents. As a result, the presence of acceptable interferents
that may have similar structures and/or properties to the targeted
solvents would not prevent the sensor from accurately reporting
existing respective solvent concentrations.
[0165] Alternatively, if the input solvent comprises certain
solvents, such as n-butanol, electrochemical oxidation could be
used to measure the solvent concentration. Electrochemical
measurements have several advantages. They are simple, sensitive,
fast, and have a wide dynamic range. The instrumentation is simple
and not affected by humidity. In one embodiment, the target
solvent, such as n-butanol, is oxidized on a platinum electrode
using cyclic voltammetry. This technique is based on varying the
applied potential at a working electrode in both the forward and
reverse directions, at a predefined scan rate, while monitoring the
current. One full cycle, a partial cycle, or a series of cycles can
be performed. While platinum is the preferred electrode material,
other electrodes, such as gold, silver, iridium, or graphite, could
be used. Although, cyclic voltammetric techniques are used, other
pulse techniques such as differential pulse voltammetry or square
wave voltammetry may increase the speed and sensitivity of
measurements.
[0166] Embodiments of the present specification expressly cover any
and all forms of automatically sampling and measuring, detecting,
and analyzing an output fluid, or the headspace above the output
fluid. For example, such automated detection can be achieved by
integrating a mini-gas chromatography (GC) measuring device that
automatically samples air in the output container, transmits it to
a GC device optimized for the specific solvents used in the
delipidation process, and, using known GC techniques, analyzes the
sample for the presence of the solvents.
[0167] The method of operation of system components 100 of FIG. 1
will now be described in detail below. FIG. 2 is a flow chart
illustrating an exemplary process for separating modified HDL, in
accordance with some embodiments of the present specification. The
method described in context of FIG. 2 may be implemented using
system components 100 described in context of FIG. 1. At 202, a
plasma delipidation process is started once the bags and tubing
sets are connected in place, as described in FIG. 1. At 204, a
first priming fluid pre-primes various fluid lines. In embodiments,
fluid lines include the tubing sets and any other channels for
transporting the fluids between the system's components.
[0168] In some embodiments, the present specification includes a
computing device with an input/output controller, at least one
communications interface and system memory. The system memory
includes at least one random access memory (RAM) and at least one
read-only memory (ROM). These elements are in communication with a
central processing unit (CPU) to enable operation of the computing
device. In various embodiments, the computing device may be a
conventional standalone computer or alternatively, the functions of
the computing device may be distributed across multiple computer
systems and architectures. In some embodiments, execution of
sequences of programmatic instructions enables or causes the
processor to perform various functions and processes. In alternate
embodiments, hard-wired circuitry may be used in place of, or in
combination with, software instructions for implementation of the
processes of systems and methods described in this specification.
Thus, the systems and methods described are not limited to any
specific combination of hardware and software.
[0169] In some configurations, the embodiments described in the
present specification include a controller having at least a
processor or processing circuitry and a system memory that is in
data communication with at least one of the basic components of the
system of the present specification to control or automate
operation of the system, including, but not limited to: [0170] One
or more fluid inputs; [0171] One or more mixing devices that may be
used to mix fluid from a fluid input and solvent from a solvent
input; [0172] One or more valves that may be used to control the
flow of a fluid, a solvent or a fluid-solvent mixture; [0173] One
or more valves that may be used to control the flow of fluid from a
fluid input and to control the flow of solvent from a solvent
input; [0174] One or more valves that may be used to control the
flow of a fluid-solvent mixture through tubing and to a separator;
[0175] One or more separators for performing bulk solvent
separation and valves associated therewith; [0176] One or more
pressure sensors and/or valves positioned in the flow stream to
control the flow of the second layer to the solvent extraction
device; [0177] One or more glucose inputs and valves associated
therewith; [0178] One or more saline inputs and valves associated
therewith; [0179] One or more solvent extraction devices; and/or
[0180] One or more pumps, which may be a peristaltic, roller, or
rotary pump.
[0181] FIG. 3A illustrates a bag 355 containing the priming fluid.
In some embodiments, the priming fluid for the first priming is
saline. Saline prepares the system by flushing the fluid flow-path
(lines) between bag 355 and a second waste container 365. The fluid
flow path flushed at this step includes a Fluid Flow Path 1 (FFP1)
comprising line 380 between bag 355 and a line 321 where line 321
is the fluid flow path between the separator and a prime connector
tube 370, a line 382 between prime connector tube 370 and the pump
350, and a line 384 extending between the pump and second waste
container 365. FFP1 is not is communication with inputs 305, 310,
360, mixer 320, first waste container 335, and output container
345. In embodiments, the pump 350 functions to draw the priming
fluid towards the prime waste or second waste container 365. In
embodiments, second waste container 365 is configured to collect
the prime waste.
[0182] In embodiments, at the preliminary priming stage, a solvent
extraction device is separate from the system. The solvent
extraction device may be a charcoal column that is subsequently
added to the system and used to extract a solvent from plasma that
contains modified HDL particles. The solvent extraction device is
substituted with prime connector tube 370 between the fluid lines
321 and 382, between bag 355 and second waste container 365.
Pre-priming the fluid lines ensures that air is substantially
removed from the fluid lines. Later when the solvent extraction
device is connected, the absence of air safeguards the function of
the solvent extraction device. In embodiments, the solvent
extraction device is a charcoal column comprising coated beads of
charcoal. The substantial or material presence of air interferes
with the efficiency and surface area of the charcoal column. In
embodiments, once the pump is closed, it does not allow for
backflow of fluids. At this stage, valves 315c, 315e, and 315g,
along FFP1 are open to facilitate passage and direct the flow of
priming fluid from bag 355 to second waste container 365 containing
the prime waste. Other valves (315, 316, 315a, 315b, 315d, 315f,
and 330) remain closed to prevent passage of the saline to other
parts of the lines, and therefore FFP1 is not in fluid
communication with inputs 305, 310, 360, mixer 320, first waste
container 335, and output container 345.
[0183] Following step 204, valves 315d and 330 are opened to
facilitate passage of priming fluid from bags 360 to separator 325,
while the other valves (315, 316, 315a, 315b, 315c, 315e, 315f, and
315g) remain closed. At 206, a priming fluid pre-primes various
fluid lines towards a separator 325. FIG. 3B illustrates at least
two bags 360 containing the priming fluid. In embodiments, the
priming fluid for priming the lines to separator 325 is saline. The
saline prepares the system by flushing the lines between bags 360
and separator 325. The fluid flow paths flushed at this step may
include line 380 between bags 360 and line 321, and line 386
extending from separator 325 to line 321. Therefore, a second fluid
flow path (FFP2) may be defined as the path including line 380
between bags 360 and line 321, and line 386 extending from
separator 325 to line 321. FFP2 does not include fluid paths to
inputs 305, 310, and 355, mixer 320, prime connector tube 370, pump
350, first waste container 335, second waste container 365, and
output container 345. The solvent extraction device is still
separate from the system.
[0184] Following step 206, valve 315d remains open, valves 315e and
315g are additionally opened, while all other valves (315, 316,
315a, 315b, 315c, 315f, and 330) remain in order to facilitate
fluid flow along a third fluid flow path (FFP3). FFP3 may be
defined as the path of fluid from bags 360, through prime connector
tube 370, to second waste container 365. FFP3 is not in fluid
communication with inputs 305, 310, and 355, mixer 320 separator
325, first waste container 335, and output container 345. At 208, a
second pre-priming operation is performed in the various fluid
lines of the system through FFP3. Referring to FIG. 3C, at this
stage, priming fluid from bags 360 are transported through valves
315d and 315e, through prime connector tube 370, and through valve
315g, toward second waste container 365, while all other valves
remain closed. Step 208 is concluded with presence of priming
fluids in the main lines of the system, which include line 380,
line 321, line 386, line 382, and line 384.
[0185] Following step 208, the priming fluid transported through
the fluid lines is drained into second waste container 365, which
is configured to collect prime waste. All the valves (315, 316,
315a, 315b, 315c, 315d, 315f, 315e, 315g, 330) are then closed.
Therefore there is no path for flow of fluids. The prime connector
tube 370 is clamped and removed from the fluid line. At 210, a
solvent extraction device is installed into the system by replacing
the prime connector tube 370. Referring to FIG. 3D, a solvent
extraction device 340 is installed at the location within the fluid
line, between lines 321 and 382, where prime connector tube 370 was
originally placed. Fluid lines (lines 321, 380, 382, 384, and 386)
between bags 355 and 360, separator 325, and second waste container
365 are pre-primed, that is, they are primed before installing
solvent extraction device 340.
[0186] Following step 210, valves 315c, 315e, and 315g are opened
while all other valves (315, 316, 315a, 315b, 330, 315d, 315f)
remain closed, which defines a fourth fluid flow path (FFP4)
extending from input 355, through solvent extraction device 340, to
second waste container 365. FFP4 is not in fluid communication with
inputs 305, 310, 360, mixer 320, separator 325, first waste
container 335, and output container 345. At 212, a first priming is
performed of the various fluid lines. Referring to FIG. 3E, priming
fluid from bag 355 is depleted and transported through FFP4
comprising valves 315c and 315e, through solvent extraction device
340, valve 315g, and into second waste container 365. The fluid
flow paths that are primed at this step include lines 380, 321,
382, and 384.
[0187] Following step 212, valve 315d is opened, valves 315e and
315g remain open, while all other valves (315, 316, 315a, 315b,
315c, 330, 315f) remain closed, which defines a fifth fluid flow
path (FFP5) extending from inputs 360, through solvent extraction
device 340, to second waste container 365. FFP5 is not in fluid
communication with inputs 305, 310, 355, mixer 320, separator 325,
first waste container 335, and output container 345. At 214, a
second priming is performed of the various fluid lines through
FFP5. Referring to FIG. 3F, priming fluid from bags 360 is
transported through valves 315d, 315e, through solvent extraction
device 340, valve 315g, and into second waste container 365. The
fluid flow paths that are primed at this step include lines 321,
380, 382, and 384.
[0188] At the conclusion of steps 212 and 214, all main fluid lines
including lines 321, 380, 382, and 384, separator 325 and solvent
extraction device 340 are primed. In embodiments, fluid lines
extending from bottom of separator 325 to second waste container
365, configured to contain prime waste, is filled with priming
fluid. Priming also results in removal of small particulates from
solvent extraction device 340.
[0189] Following step 214, valve 315 is opened and all other valves
(316, 315a, 315b, 315c, 315d, 315e, 315f, 315g, and 330) are
closed, defining a sixth fluid flow path (FFP6) from input 305 to
mixer 320. FFP6 is not in fluid communication with bag, 310,
separator 325, first waste container 335, inputs 310, 355, 360,
solvent extraction device 340, pump 350, second waste container
365, and output container 345. At 216, the plasma fluid and the
solvent are introduced one after the other, into a mixing device of
the system. In embodiments, a blood fraction of the patient is
obtained, which in a still further embodiment is plasma. The
process of blood fractionation is typically achieved by filtration,
centrifuging the blood, aspiration, or any other method known to
persons skilled in the art. Blood fractionation separates the
plasma from the blood. In an embodiment, blood fractionation is
performed remotely. In one embodiment, blood is withdrawn from a
patient in a volume sufficient to produce about 12 ml/kg of plasma
based on body weight. During the fractionation process, the blood
can optionally be combined with an anticoagulant, such as sodium
citrate, and centrifuged at forces approximately equal to 2,000
times gravity. The blood is separated into plasma and red blood
cells using methods commonly known to one of skill in the art, such
as plasmapheresis. In an embodiment, the red blood cells are then
aspirated from the plasma. In one embodiment, the process of blood
fractionation is performed by withdrawing blood from the patient
with the cardiovascular and/or related disease, and who is being
treated by the physician. In an alternative embodiment, the process
of blood fractionation is performed by withdrawing blood from a
person other than the patient with the cardiovascular and/or
related disease who is treated by the physician. Therefore, the
plasma obtained as a result of the blood fractionation process may
be either autologous or non-autologous.
[0190] Subsequent to fractionation, the red blood cells are either
stored in an appropriate storage solution or, preferably, returned
to the patient during plasmapheresis. Physiological saline, 5%
albumin, or other suitable fluid may also optionally be
administered to the patient to replenish volume. If the blood was
obtained from an individual other than the patient, the cells are
returned to that individual, who can also be referred to as the
donor.
[0191] Plasma obtained from blood is usually a straw-colored liquid
that comprises the extracellular matrix of blood cells. Plasma is
typically 95% water, and contains dissolved proteins, which
constitute about 6-8% of plasma. The plasma also contains glucose,
clotting factors, electrolytes, hormones, carbon dioxide, and
oxygen. The plasma has a density of approximately 1006 kg/m3, or
1.006 g/ml.
[0192] In some alternate embodiments, Low Density Lipoprotein (LDL)
is also separated from the plasma. Separated LDL is usually
discarded. In alternative embodiments, LDL is retained in the
plasma. In accordance with embodiments of the present
specification, blood fraction or plasma obtained includes plasma
with High Density Lipoprotein (HDL), and may or may not include
other protein particles. In embodiments, autologous or
non-autologous plasma collected from the patient or donor,
respectively, is subsequently treated via an approved
plasmapheresis device. The plasma may be transported using a
continuous or batch process.
[0193] Referring to FIG. 3G, a plasma input bag 305 contains the
plasma that will be treated by the various embodiments of the
present specification. The plasma is transported from bag 305,
along FFP6, through a valve 315, into a mixing device 320.
[0194] Following transportation of plasma from bag 305 to mixing
device 320, valve 315 is closed and valve 316 is opened, while all
the other valves (315a, 315b, 315c, 315d, 315e, 315f, 315g, 330)
remain closed, thus defining a seventh fluid flow path (FFP7) from
bag 310 to mixing device 320. FFP7 is not in fluid communication
with bag 305, separator 325, first waste container 335, inputs 310,
355, 360, solvent extraction device 340, pump 350, second waste
container 365, and output container 345. Referring to FIG. 3H, a
solvent input bag 310 contains the solvent. The solvent is
transported along FFP7, from bag 310, through a valve 316, into
mixing device 320. The solvent is used for extracting lipids from
the plasma fluid or from particles within the plasma fluid.
Suitable extraction solvents include solvents that extract or
dissolve lipid, including but not limited to phenols, hydrocarbons,
amines, ethers, esters, alcohols, halohydrocarbons, halocarbons,
and combinations thereof. Examples of suitable extraction solvents
are ethers, esters, alcohols, halohydrocarbons, or halocarbons
which include, but are not limited to di-isopropyl ether (DIPE),
which is also referred to as isopropyl ether, diethyl ether (DEE),
which is also referred to as ethyl ether, lower order alcohols such
as butanol, especially n-butanol, ethyl acetate, dichloromethane,
chloroform, isoflurane, sevoflurane (1,1, 1,3,
3,3-hexafluoro-2-(fluoromethoxy) propane-d3),
perfluorocyclohexanes, trifluoroethane, cyclofluorohexanol, and
combinations thereof. In an embodiment, a mix of Sevoflurane and
n-butanol is used as the solvent. In an embodiment a volume ratio
of Sevoflurane and n-butanol used is 95:5. In various embodiments,
the plasma and the solvent are transported in any order. While
transporting the plasma and the solvent to mixing device 320, the
valves corresponding to bags containing the plasma (valve 315) and
the solvent (valve 316) are respectively open. All other valves
(315a, 315b, 315c, 315d, 315e, 315f, 315g, and 330) remain
closed.
[0195] Following step 216, once the plasma and the solvent are in
mixing device 320, all the valves (316, 316, 315a, 315b, 315c,
315d, 315e, 315f, 315g, 330) are closed. At 218, the plasma and the
solvent, which are present together within the mixing apparatus are
processed by a mixing operation. In an embodiment, the solvents
used include either or both of organic solvents sevoflurane and
n-butanol. In embodiments, the solvent is optimally designed such
that only the HDL particles are treated to reduce their lipid
levels and LDL levels are not affected.
[0196] The mixing process includes factoring in variables such as
the solvent employed, mixing method, time, and temperature. Choice
of a mixing method may also affect some system requirements, such
as but not limited to functional requirements, packaging
requirements, cost, and environmental requirements. Additional
variables that may be considered in system design are as follows:
[0197] 1. Priming. Whether the system can handle priming so that it
can be primed and ready for a process. [0198] 2. Continuous or
Interrupted Flow. Whether the system has to be continuous in nature
(continuously separates plasma and inputs replacement fluid in
parallel) or can handle discrete amounts of liquid (batch flow).
[0199] 3. Closed Loop Control. Whether the system has to be
monitored or can be validated by process. [0200] 4. Variable Flow.
Whether the system can handle various ranges of flow. [0201] 5.
Hold-up Volume. This is the amount of fluid or blood that remains
in the circuit after the process is complete, as it is desirable to
have as little blood or fluid outside of the patient at any given
time and so that the plasma that will be returned to the patient is
not overdiluted. [0202] 6. Mixing Control. Whether the system
allows for controlling the level, speed or extent of mixing. [0203]
7. Plasma Range. Whether the system can handle different types of
plasma, including normal plasma, high LDL plasma, high triglyceride
plasma, and other types of plasma. [0204] 8. Packaging
Requirements. Whether a hospital or blood bank could accommodate
the footprint of the system and its associated components,
including disposables and hardware. [0205] 9. Environmental
Requirements. Whether the system could be deployed in a variety of
settings with respect to operating noise/vibration and hardware
durability (for example, whether it can be deployed in bloodbanks
or hospitals). [0206] 10. Cost. Whether the system can be
manufactured in a cost-effective manner (for example, no high cost,
high precision connectors).
[0207] Different mixing methods provided different results in terms
of remaining cholesterol, remaining phospholipids, remaining Apo-B,
and remaining Apo-A, within the delipidated plasma obtained after
the mixing. The mixing methods employed in the present
specification take into account a plurality of variables that, when
combined, achieve an ideal mixing environment for optimal selective
delipidation. By way of background, several mixing methods, as are
well-known to those of ordinary skill in the art were initially
employed with little to no success. These methods included
continuous vortex mixing, mixing using a static mixer, mixing using
a silly straw mixer, and mixing using a rotating cylinder.
[0208] Continuous vortex type of mixing involves using a vortexer
to mix smaller quantities of liquid. When a test tube or other
container is pressed into the rubber cup of the vortexer, the
motion is transferred to the liquid inside, creating a fluid vortex
or whirlpool in an off-center rotation. Because the speeds achieved
are close to 2500 rpm, the end result could be "overdelipidation",
or complete delipidation of both HDL and LDL. As discussed above,
it is not desirable to delipidate LDL.
[0209] A static mixer is a plate-type mixer or a mixer comprising
mixing elements contained within an elongated housing that
effectuates movement of a tube containing a mixture or mixture of
fluids, where the movement is typically sideways from one side to
another. It is typically employed for continuous mixing. This
method of mixing does not work as it 1) involves direct connection
to the patient for serial apheresis and delipidation and also
results in "overdelipidation", or complete delipidation of both HDL
and LDL. As discussed above, it is not desirable to delipidate
LDL
[0210] The "silly straw" method of mixing was designed as a coiled
tube (a tubing set wrapped around a stick) through which a fluid
mixture flows continuously creating a Taylor vortex. A continuous
flow of plasma and solvent in a 2:1 solvent to plasma ratio was
used. This method proved to be entirely ineffective. One theory is
that in order to effectively selectively delipidate, the plasma and
solvent mixture needs to be in full contact at a specified ratio,
for a specified amount of time and that an instantaneous
flow-through process could not achieve this equilibrium.
[0211] Mixing using a rotating cylinder involves a tube containing
the mixture that rotates around its axis (similar to the movement
of a record on a turntable) to mix the fluids within the tube. In
using this process, the fluid tumbles from the top to the bottom of
the test tube. While the method was not optimal for selective
delipidation as described in the present specification, a novel
mixing bag of specific geometry and size was designed and
implemented.
[0212] The system of the present specification, and in particular,
the mixing sub-system is designed so that it can be primed and
ready for a process. In addition, the system of the current
specification can handle fluid processing in batches without the
need for continuous flow. The system of the present specification
advantageously does not require closed loop control. Once the
parameters (flow rate, volume) are established, the system is
gravity based and operates accordingly. Optionally, a charcoal
column is employed to further ensure that all solvent is removed.
The system of the present specification can also accommodate
various ranges of fluid flow. Because the system of the present
specification is a stand-alone system (meaning that apheresis is
not integrated), the issue of hold-up volume becomes a non-issue.
The system of the present specification also allows for controlling
the level of mixing by determining the speed of the mixer and using
a mixing bag with an appropriate volume and geometry. Further, the
system of the present specification is designed to be able to
handle a wide and infinite range of plasma that can be treated by
the system, including, but not limited to normal plasma, high LDL
plasma, high triglyceride plasma, and other types of plasma. The
system of the present specification has low to minimal footprint,
is readily and easily deployable in a variety of environments with
minimal noise impact. In addition, the system of the present
specification can be manufactured in a cost-effective manner.
[0213] Referring to FIG. 3I, mixing device 320 may be a bag used
for mixing the plasma and the solvent. In embodiments, mixing
device 320 includes both an orbital mixer and a mixing bag, and is
placed at an angle within the system. In one embodiment, the mixing
bag is placed horizontally over the orbital mixing device, because
this position may receive the most optimal orbital mixing action
for the fluid contained in the bag, as most of the fluid would
gravitate towards the bottom. The bag and its corners can be used
to impart energy. The mixing bag, using an angled platform as
described below may be placed at a slight angle to enable draining
of the fluids. The angle may range from 0 degrees (completely
vertical) to 90 degrees (completely horizontal). In one embodiment,
the platform upon which the mixing bag rests is placed at an angle
of 18.2 degrees. In one embodiment, mixing device 320 is of a
circular shape. In another embodiment, mixing device 320 is of a
rectangular shape.
[0214] FIG. 7 illustrates an exemplary mixing device (bag) 700, in
accordance with embodiments described in context of FIG. 3I. Bag
700 is of a rectangular shape, comprising a section 702 where
plasma and solvent fluid may be received through an input pipe 708.
Section 702 has at least five edges and is at the center of bag
700. Section 702 is surrounded by sealed sections of bag 700. One
of the edges of bag 700 includes a rectangular sealed section 704.
Two mutually inclined edges, opposite to the edge along section
702, include triangular sealed sections 706a and 706b. Each sealed
section 704, 706a, and 706b, includes at least one hanger hole,
such as holes 712. Section 702 includes an output pipe 714
positioned between triangular sealed section 706a and 706b, to
enable letting out of the mixture.
[0215] FIG. 8A illustrates a side view of shaker angle brackets 800
that are used to position mixing device 320 within the system, in
accordance with some embodiments of the present specification. FIG.
8B illustrates a another side view of shaker angle brackets 800
that are used to position mixing device 320 within the system, in
accordance with some embodiments of the present specification. FIG.
8C illustrates a perspective view of shaker angle brackets 800 that
are used to position mixing device 320 within the system, in
accordance with some embodiments of the present specification.
Referring simultaneously to FIGS. 8A, 8B, and 8C, the mixing bag
used to perform the mixing operation may be placed over an orbital
mixer, which is fitted within the system with the help of brackets
800. In embodiments, brackets 800 are manufactured from Aluminum.
In one embodiment, the Aluminum used for making brackets 800 is
0.060 inches thick. Referring simultaneously to FIGS. 8A, 8B, and
8C, brackets 800 include two parts 802 and 804, which mirror each
other. In one embodiment, bracket 802 is placed on the left and
bracket 804 is placed opposite to bracket 802 on the right. A
mixing device, such as device 320 of FIG. 3I, is positioned on
brackets 802 and 804. Holes 806 on both brackets 802 and 804 enable
fixing the mixing device. In one embodiment, the mixing device
provides a platform for placing the mixing bag, such as bag 700 of
FIG. 7. Each bracket has two opposing sides connected by a flat
surface between them. A top side 808 of each bracket is inclined at
an angle for placing the mixing bag. In embodiments, the angle is
in the range of 0 to 90 degrees. In one embodiment, the angle is
18.2 degrees, relative to a horizontal bottom side 810. The incline
provided by top side 808 enables placing the mixing device, and
therefore the mixing bag at an inclination, for an optimal mixing
operation. Each edge 808 and 810 is bent in two ways to created
angled brackets 802 and 804. The bent portions include holes 806
that enable fixing the mixing device with brackets 800.
[0216] In one embodiment, mixing device 320 has a capacity of 300
milliliters (ml). In one embodiment, mixing device 320 is
configured to mix approximately 100 ml of plasma with the solvent
during a single mixing operation. In one embodiment, solvent and
plasma are mixed in a volume ratio of 2:1. For example, 100 ml of
plasma is mixed with 200 ml of the solvent. In another embodiment,
the solvent and plasma are mixed in a volume ratio of 1:1.
[0217] Solvent type, ratios and concentrations may vary in this
step. Acceptable ratios of solvent to plasma include any
combination of solvent and plasma. In some embodiments, (volume)
ratios used are 2 parts plasma to 1 part solvent, 1 part plasma to
1 part solvent, or 1 part plasma to 2 parts solvent. In an
embodiment, when using a solvent comprising 95 parts sevoflurane to
5 parts n-butanol, a ratio of two parts solvent per one part plasma
is used. Additionally, in an embodiment employing a solvent
containing n-butanol, the present specification uses a ratio of
solvent to plasma that yields at least 5% n-butanol in the final
solvent/plasma mixture. In an embodiment, a final concentration of
n-butanol in the final solvent/plasma mixture is 3.33%. In
embodiments, the final concentration of n-butanol in the resultant
solvent/plasma mixture may vary and may be dependent on the solvent
to plasma ratio, which may also vary. The plasma may be transported
to the mixing device using a continuous or batch process. Further
various sensing means may be included to monitor pressures,
temperatures, flow rates, solvent levels, and the like. The
solvents dissolve lipids from the plasma. In embodiments of the
present specification, the solvents dissolve lipids to yield
treated plasma that contains modified HDL particles with reduced
lipid content. The process is designed such that HDL particles are
treated to reduce their lipid levels and yield modified HDL
particles without destruction of plasma proteins or substantially
affecting LDL particles. It should be noted that there is no
clinically significant decrease in blood constituents
post-plasmapheresis.
[0218] In one embodiment, mixing device 320 is operated to mix the
plasma solvent mixture for 60 seconds with an average mixing plasma
batch volume of 99.+-.7.5 ml.
[0219] In various embodiments, various energy measurements are
provided as input to operate mixing device 320. Energy is
introduced into the system in the form of varied mixing methods,
time, and speed. A combination of the mixing parameters such as but
not limited to the volume ratio of solvent to plasma, shape of the
mixing device 320, and the amount of energy input used to operate
mixing device 320, directly affect the success of the mixing
operation to achieve delipidated HDL particles in the plasma. In
one example, a solvent to plasma ratio of 2:1, for a batch of 100
ml plasma, mixed for 60 seconds, in a rectangular mixing device,
using an energy input of 200 RPM does not delipidate the HDL
particles from the plasma. In another example, a solvent to plasma
ratio of 2:1, for a batch of 100 ml plasma, mixed for 60 seconds,
in a large square mixing device, using an energy input of 400 RPM
also does not delipidate the HDL particles from the plasma.
Therefore, multiple parameters affect the success of delipidating
HDL particles from the plasma. The effect of varying the different
parameters is described in the subsequent sections, and in context
of experiments illustrated in FIGS. 4, 5, and 6.
[0220] Referring back to step 218 of FIG. 2, and FIG. 3I, the
plasma and the solvent interact with each other within mixing
device 320 to the extent that HDL particles are delipidated, while
LDL particles are not. The process is therefore termed as selective
delipidation. In embodiments, the mixing is performed in order to
achieve at least 80% delipidation of HDL particles.
[0221] After the mixing, valve 315a is opened while all the other
valves (315, 316, 315b, 315c, 315d, 315e, 315f, 315g, 330) remain
closed, thus defining an eighth fluid flow path (FFP8) between
mixing device 320 and separator 325. FFP8 is not in fluid
communication with bags 305, 310, 355, 360, waste containers 335,
365, solvent extraction device 340, pump 350, and output container
345. At 220, once the mixing operation is completed, the solvent
plasma mixture is transferred to a separator along FFP8, where the
plasma and the solvent are separated by gravity. Referring to FIG.
3J, mixture of solvent and plasma is dropped down through a valve
315a in to a separator 325. The mixture remains in separator 325
until the solvent settles at the bottom of separator 325. The
plasma is separated and remains in a layer above the solvent. The
solvent employed is preferably of a higher density than plasma, and
therefore settles at the bottom.
[0222] In embodiments, steps 216, 218, and 220 are performed
iteratively, in batches, until separator 325 is filled to its
capacity. Once the separator is filled to its capacity, all the
valves (315, 316, 315a, 315b, 315c, 315d, 315e, 315f, 315g, 330)
are closed. At 222, and referring to FIG. 3K, the collected mixture
of plasma and solvent in separator 325 is allowed to stand for a
period of time, until the solvent separates and settles at the
bottom of separator 325. In embodiments, the period of time is
dependent upon the time it takes for the solvent to fully separate
without a loss or sacrifice in the amount of plasma. In one
embodiment, the mixture is allowed to stand for approximately 30
minutes. In embodiments, separator 325 has a cone-shaped bottom
that enables easy removal of bulk solvent in a subsequent step.
[0223] Following the separation, valve 315b is opened to define a
ninth fluid flow path (FFP9) from separator 325 to first waste
container 335. FFP9 is not in fluid communication with bags 305,
310, 355, 360, mixer 320, second waste container 365, solvent
extraction device 340, pump 350, and output container 345. At 224,
bulk solvent is removed from the separator 325 along FFP9.
Referring to FIG. 3L, bulk solvent that has settled at the bottom
portion of separator 325 flows to a first waste container 335
through a valve 315b, which, when open allows fluid to flow freely
using gravity. In other embodiments, a pump may be employed to
remove the solvent. Cone-shaped bottom of separator 325 aids easy
removal of bulk solvent. Valve 315b is closed after bulk solvent
has been moved through it and before the plasma from separator 325
reaches valve 315b.
[0224] In embodiments, a weight of separator 325 is known, in
addition to weight of the plasma and of the solvent. In
embodiments, the weight of the separator bag is continuously
monitored. With this information, valve 315b is closed as soon as
the amount of solvent removed from separator 325 corresponds to the
known weight of the solvent. The weight of the solvent that flows
to the first waste container 335 is, in an embodiment, indirectly
monitored, because the amount of solvent that is added to the
system and the amount of solvent present in the separator waste bag
are known. In addition, the residual concentration of solvent that
is in the plasma is based on validation of system parameters and a
validated analysis of residual solvent concentrations via GC over
many process runs.
[0225] Once valve 315b is closed, valves 330, 315e, and 315g are
opened while all other valves (315, 316, 315a, 315c, 315d, 315f,
330) remain closed, thus defining a tenth fluid flow path (FFP10).
FFP10 is not in fluid communication with bags 305, 310, 355, 360,
mixing device 320, first waste container 335, second waste
container 365, and output device 345. At 226, and referring to FIG.
3M, a pump 350 is turned on and valves 330 and 315e are opened in
order to pull plasma from separator 325 through fluid line 321
towards solvent extraction device 340, along FFP10. During this
operation, initially a valve 315g is simultaneously open. Valve
315g is placed between solvent extraction device and second waste
container 365. As the pump pulls the fluid present in lines 321
from separator 325 through solvent extraction device 340, priming
fluid that was initially present in lines 321, 382, and 384,
extending between separator 325 and second waste container 365, is
pushed, or chased, further ahead in the lines by the plasma being
pulled through valve 315g, towards second waste container 365
configured to contain prime waste. Once plasma (pulled by pump 350)
reaches valve 315g, which is determined using both the tube length
and the volume of fluid passed through the pump per revolution, the
valve 315g is closed so that priming fluid is separated from
plasma. This ensures that the plasma is not diluted and additional
fluids are not collected along with the plasma that will
subsequently be delivered back to the patient.
[0226] Subsequently, valve 315f is opened in addition to already
open valves 330, 315e, while all other valves (315, 316, 315a,
315b, 315c, 315d, 315g) remain closed, thus defining another fluid
flow path (FFP11) from separator 325, through solvent extraction
device 340, to output device 345. At 228, and referring to FIG. 3N,
pump 350 further pulls plasma along FFP11, from separator 325
through valves 330 and 315e, through solvent extraction device 340,
and through a valve 315f, towards and into an output plasma
container 345. As plasma moves through solvent extraction device
340, charcoal in solvent extraction device 340 absorbs and
therefore extracts any remaining solvent from the plasma.
[0227] After extracting the delipidated plasma in to output
container 345, valve 330 is closed along with valves 315, 316,
315a, 315b, 315c, 315g, and valve 315d is opened along with open
valves 315e and 315f, thus defining another fluid flow path FFP12
from bags 360 through solvent extraction device 340, to output
device 345. FFP 12 is not in fluid communication with bags 305,
310, 355, mixing device 320, separator 325, first waste container
335, and second waste container 365. At 230, and referring to FIG.
3O, once plasma is pulled out from separator 325 completely, pump
350 is still operated until priming fluid from bags 360 along FFP12
to follow or chase the plasma in the lines 380, 321, 382, and 384,
through valve 315e, through solvent extraction device 340, and
through valve 315f. Pump 350 is stopped once the priming fluid,
chasing the plasma, reaches a position in the fluid line which is
just before reaching output plasma container 345. In an embodiment,
150 mL of priming fluid is used to further chase the plasma into
the plasma output bag 145/345 to ensure full recovery of the
delipidated plasma. In an embodiment, chasing the fluid flow to the
prime waste occurs to the point where pump 350 reaches a specific
number of revolutions that is indicative of the plasma volume that
has flowed through the system. Thus, the revolutions of the pump
control how much fluid is in the prime waste or second waste
container 345. Pump 350 is stopped to ensure that as much of the
delipidated plasma that available in the system is collected in
container 345, while the collected plasma is saved from unnecessary
dilution by priming fluids. In embodiments, pump 350 is stopped
automatically by the system based on the amount of plasma
collected, which corresponds to the known amount of input plasma.
In embodiments, the configurations of the disposable elements
within the system are employed to program the system to
automatically stop pump 350. In an embodiment, the tubing sets are
of a known length and diameter. In addition, the volume of solvent
at the bottom of separator bag 325 is also known in addition to the
amount of plasma.
[0228] The extracted modified HDL plasma solution has an increased
concentration of pre-beta HDL. It is estimated that the modified
HDL in the delipidated plasma, has approximately 80-85% of
pre-.beta. particles, and about 15% of .alpha. HDL particles.
Concentration of pre-beta HDL is greater in the modified HDL,
relative to the original HDL that was present in the plasma before
treating it with the solvent. Compared to the plasma solution
originally separated from the blood fraction, which typically
contains approximately 5% of pre-.beta. HDL particles, the
concentration of pre-.beta. HDL particles is substantially
increased.
[0229] At the end of this process, solvent waste is collected
separately in first waste container 135/335, and prime waste is
collected in second waste container 165/365, through their separate
waste streams. This is advantageous for many reasons. Primarily, it
is more expensive to dispose of certain types of waste, such as
solvent waste. If solvent waste is "contaminated" with or combined
with other types of waste, the additional waste will have to be
disposed of in the same costly manner as solvent waste. Prime
waste, for example, which consists of mostly saline and/or glucose,
can be directed to a normal hospital waste stream. If mixed with
solvent, the prime waste will have to be diverted into the chemical
waste disposal channels, by default. By separating out waste, each
waste stream can be treated and disposed of appropriately. In some
embodiments, the solvent waste can be treated or scrubbed to
reclaim a pure solvent so that it can be re-used.
[0230] Examples of effect of varying multiple parameters are now
explained briefly. Among various methods by which plasma may be
delipidated, parameters affecting the extent of delipidation may be
broadly identified as chemical and mechanical parameters. Examples
of chemical parameters may include, but may not be limited to,
plasma type (bovine, human, lipemic), plasma volumes, solvent type
(n-butanol/DiPE or n-butanol/sevoflurane, any other), percentage of
n-butanol present in the solvent, and solvent to plasma ratio.
Examples of some of the mechanical parameters may include, a method
of mixing (rocker table, vortex, any other), mixing duration,
method of separation (gravity, centrifuge, any other), separation
time, and centrifuge force.
[0231] For purposes of illustrating the effect of varying these
parameters on the extent of delipidation, an experimental
delipidation process was performed in a laboratory setting. The
results, presented in table 400 of FIG. 4, are briefly discussed
herein. Referring to table 400, first column 402 lists different
embodiments in separate rows. Each embodiment corresponds to a
unique combination of the parameters that affect the extent of
delipidation. The second column 404 lists the plasma type used for
each embodiment. The plasma type was selected from the plasma of a
human or that of a bovine. Column 406 lists the plasma volumes (in
milliliter) used in each embodiment. Column 408 lists the type of
solvent used. In most embodiments, the solvent type is either of
n-Butanol and DiPe. Column 410 lists the percentage of n-Butanol
used, which may also be inferred as an indication of the solvent
ratio. Column 412 lists the solvent to plasma ratio used in each
embodiment. Column 414 lists the type of mixing method used for
each embodiment. Column 416 lists the time for which the mixing
process was implemented. Column 418 lists the chosen method for
separation of the plasma and the solvent. Column 420 lists the time
for which the process of separation was performed for each
embodiment. Column 422 lists the amount of centrifugal force
applied for each embodiment. Lastly, column 424 lists the results
that show the variation across each embodiment, in the percentage
of lipids that remain in the treated plasma.
[0232] Embodiment 1: About 10 milliliters (ml) of plasma derived
from a human was used. This plasma was mixed with n-butanol/DiPE
solvent. A solvent to plasma ratio of 2:1 was used. The rocker
table was used to perform the mixing operation, for about five
minutes. A centrifugal force of 563.times.G was applied for about
two minutes to separate the delipidated plasma from the solvent.
The effect of varying percentage of n-butanol in the solvent within
a range of 0% to 40% is that remaining lipids progressively
decrease with an increase in the quantity of n-butanol in the
solvent.
[0233] Embodiment 2: In another similar experiment, 10 ml of bovine
plasma was mixed with n-butanol/DiPE solvent containing 25%
n-butanol. The mixture was mixed using a rocker table for about
five minutes. A centrifugal force of 563.times.G was applied for
about two minutes to separate the delipidated plasma from the
solvent. The effect of varying the solvent to plasma ratio in a
range of 0.25 to 10 is that a lower ratio, specifically within a
range of 1 to 2, results in most reduced lipid concentration in the
delipidated solution.
[0234] Embodiment 3: In another similar experiment, 10 ml of human
plasma was mixed with n-butanol/DiPE solvent containing 25%
n-butanol, using a solvent to plasma ratio of 2:1. Different
samples of the mixture were mixed using a rocker table and using a
vortex. Gravity separation was used for about five minutes to
separate some of the samples, as well as a centrifugal force of
563.times.G was applied for about two minutes to separate the
delipidated plasma from the solvent for the remaining samples. The
effect of different mixing methods and by varying the duration of
mixing for both the methods used for separating (gravity and
centrifuge) is that there is a variation on the concentration of
lipids remaining in the delipidated plasma.
[0235] Embodiment 4: In yet another similar experiment, 10 ml of
human plasma was mixed with n-butanol/DiPE solvent containing 25%
n-butanol, using a solvent to plasma ratio of 2:1. The mixture was
mixed using a rocker table for about five minutes. A range of
centrifugal force was applied for about two minutes to separate the
delipidated plasma from the solvent. The effect of varying the
centrifugal force used for separation on the lipid concentration
remaining in the delipidated plasma
[0236] FIG. 5 is a table 500 that lists another exemplary set of
variables that may affect the delipidation process and resultant
percentage selective delipidation and is presented by way of
example only to show possible combinations of variables. The ideal
results from these experiments include a substantial change in HDL
concentration, no change in LDL concentration, preservation of
Apo-A1, preservation of Apo-B, and preservation of phospholipids,
resulting in selective delipidation of plasma. Referring to the
table 500, the first column 502 lists the type of solvent mix used.
The constituents for the solvent solution may contain one or more
of Sevoflurane (S), n-butanol (N), DiPE (D), and Isofluorane (I).
The second column 504 (Solvent Ratio) lists the ratio of the
constituents of the solvent that may be used in the solvent
solution, corresponding to the first column. The third column 506
(Plasma:Solvent Ratio) lists the proportion of the plasma and the
solvent that may be mixed together for the delipidation. The fourth
column (Mix Method) 508 states the corresponding mixing method that
may be used. The fifth column (Time) 510 provides the corresponding
duration for which mixing can be performed. The sixth column (Sep.
Method) 512 lists the method used for separation of the delipidated
plasma and the solvent. The two methods commonly used for
separation are gravity separation (GS) and centrifugal separation
(CF), in the embodiments of the present specification.
[0237] FIG. 6 is a table 600 that provides another exemplary set of
variables that may be used for normal plasma and lipemic IV plasma
using different solvents and different methods of separation. A
first column 602 (Plasma) lists the type of plasma (normal or
lipemic IV) used in each embodiment, where each row corresponds to
a different embodiment. Column 604 (solvents) lists the type of
solvent or solvent mixture used corresponding to each embodiment.
Column 606 (Ratio) lists the ratios of constituents in a solvent
mixture for each embodiment. Column 608 (P:S) lists the plasma to
solvent ratio corresponding to each embodiment. Column 610 (Volume)
lists the volume of the plasma used for each embodiment. Column 612
(S Volume) lists the volume of the solvent/solvent mixture used for
each embodiment. The volumes of the plasma and the solvent/solvent
mixture corresponds to the ratio listed in column 608. Column 614
(Mix Method) lists the method of mixing used for each embodiment.
Column 616 (Time) lists the duration for which the mixing was
performed. Column 618 (Separation) lists the method used for
separation (centrifugal or gravity separation) of the plasma and
the solvent. Column 620 (time(min)) lists the duration (in minutes)
for which the separation process was performed for each embodiment.
Lastly, column 622 (Solvent Removal) lists the type of method used
for solvent removal. As seen in table 600, a charcoal column was
used in all the embodiments to remove the solvent.
[0238] In general, the present specification preferably comprises
configurations wherein all inputs, such as input plasma and input
solvents, disposable elements, such as mixing bags, separator bags,
waste bags, solvent extraction devices, and solvent detection
devices, and output containers are in easily accessible positions
and can be readily removed and replaced by a technician.
[0239] To enable the operation of the above described embodiments
of the present invention, it is preferable to supply a user of such
embodiments with a packaged set of components, in kit form,
comprising each component required to practice embodiments of the
present specification. The kit may include an input fluid container
(i.e. a high density lipoprotein source container), a lipid
removing agent source container (i.e. a solvent container),
disposable components of a mixer, such as a bag or other container,
disposable components of a separator, such as a bag or other
container, disposable components of a solvent extraction device
(i.e. a charcoal column), an output container, disposable
components of a waste container, such as a bag or other container,
solvent detection devices, and, a plurality of tubing and a
plurality of valves for controlling the flow of input fluid (high
density lipoprotein) from the input container and lipid removing
agent (solvent) from the solvent container to the mixer, for
controlling the flow of the mixture of lipid removing agent, lipid,
and particle derivative to the separator, for controlling the flow
of lipid and lipid removing agent to a waste container, for
controlling the flow of residual lipid removing agent, residual
lipid, and particle derivative to the extraction device, and for
controlling the flow of particle derivative to the output
container.
[0240] In one embodiment, a kit comprises a plastic container
having disposable components of a mixer, such as a bag or other
container, disposable components of a separator, such as a bag or
other container, disposable components of a waste container, such
as a bag or other container, and, a plurality of tubing and a
plurality of valves for controlling the flow of input fluid (high
density lipoprotein) from the input container and lipid removing
agent (solvent) from the solvent container to the mixer, for
controlling the flow of the mixture of lipid removing agent, lipid,
and particle derivative to the separator, for controlling the flow
of lipid and lipid removing agent to a waste container, for
controlling the flow of residual lipid removing agent, residual
lipid, and particle derivative to the extraction device, and for
controlling the flow of particle derivative to the output
container. Disposable components of a solvent extraction device
(i.e. a charcoal column), the input fluid, the input solvent, and
solvent extraction devices may be provided separately.
[0241] The above examples are merely illustrative of the many
applications of the system of present invention. Although only a
few embodiments of the present invention have been described
herein, it should be understood that the present invention might be
embodied in many other specific forms without departing from the
spirit or scope of the invention. Therefore, the present examples
and embodiments are to be considered as illustrative and not
restrictive, and the invention may be modified within the scope of
the appended claims.
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