KR880001097B1 - Biospecific polymer - Google Patents

Abstract

Biocompatible polymers are prepd. by immobilizing reactive biol. agents on a polymer support via. chem. bonding. Thus, solns. of monomer are prepd. by combining 2-hydroxyethyl methacrylate, ethylene glycol, NaHSO3 and ammonium persulfate. After polymn., polymer membrane disks are cut out from the polymer sheet. The disks are activated in 10% (NBr soln. Antibodies are obtd. by incubating poly (2-hydroxyethyl methacrylate) membrane disks which are treated with insulin-absorbed anti-insulin antibody. These insulin-treated membrane disks absorbed 111 pg antiinsulin antibody from soln. or 690 pg/cm2 surface area in a radioimmuno assay test.

Description

Biospecific Polymer

FIELD OF THE INVENTION The present invention relates to biocompatible polymers, which are affixed with fixed reactive biochemicals to selectively bind certain pathogens or populations thereof associated with fluids of a diseased organism.

Various progressions of vaginal manifestations often result in elevated levels of certain blood proteins and other molecules. This phenomenon is usually used as a diagnostic method to define the condition and to check the course of treatment. In many cases, this particular blood component is a direct or indirect cause of primary and secondary signs of disease progression. Autoimmune diseases can be characterized by the circulation of antibodies in endogenous substrates and tissue protein gels necessary for normal growth and maintenance of the living body. Neoplastic diseases are characterized by the growth of an unmodified cell line that escapes or damages the natural defenses of the body by generating immunosuppressive blockers, surface antigen masking components and / or growth regulators. Selective separation of these pathogens on a biocompatible substrate is part of restoring normal function of the body by eliminating pathologies of disease progression.

The fundamental function of organs, cells and molecules, including the immune system, is to recognize and remove foreign substances from the living body. These foreign substances are removed by working with the antibodies formed in response to that substance. In general, this function is effectively performed without harming the patient. In some cases, however, disturbances may occur, leading to pathogenic diseases such as, for example, an uncontrolled reaction (allergic disease) or an abnormal reaction (autoimmune disease). Hospitals of son's disease are directly or indirectly involved in the production of antibodies with heterologous reactivity with environmental antigens (allergens) or autoantigens.

Autoimmune diseases are pathologies that occur when a patient reacts immunologically by the production of antibodies that are reactive with autoantigens. Autoimmunity can affect almost any part of the body and generally involves a reaction between autoantigen and immunoglobulin (IgM or IgG) antibodies. Representative autoimmune diseases may include the thyroid gland, kidney, pancreas, nerve cells, gastrointestinal mucosa, adrenal gland, skin, red cells, synovial membrane, thyroid hemoglobin, insulin, deoxyribonucleic acid, immune hemoglobin, and the like.

For some forms of autoimmune and neoplastic diseases, limited success can be achieved by non-selective immunosuppressive treatment, such as by projecting x-rays or administering cytotoxic agents throughout the body. Drawbacks of these treatments include the toxicity of the agents used and the development of various tumors, particularly lymphoid and gastric cell tumors. In addition, the use of unspecified drugs for long-term cell suppression increases the likelihood that patients will be severely infected with fungi, bacteria, and viruses that would not normally be a problem. The present invention is optional in the sense of removing only the causative pathogen or its population in association with the manifestation of a particular disease.

There are generally two conventional methods of treating autoimmune and / or neoplastic diseases. One way is to inject a patient with a substance that produces a specific form of immunological resistance. Inhibition of the antibody reaction in this way results in resistance to the antigen against it. Representative examples include therapies in which the conjugate of the antigen bound to D-glutamic acid, ie, the D-lysine copolymer, is injected into the patient.

Another treatment is to use routes outside the body. This method usually involves the removal of whole blood, the separation of cytosolic and lysable blood substances, the substitution or treatment of a tablet and the recombination-infusion of the treated whole blood. One example of this method is to replace or exchange plasma with salt, sugar and / or protein solutions. However, plasma exchange is a crude technique and requires a large amount of replacement solution. Another example is the physical and / or biochemical alteration of the plasma portion of the whole blood. There are several specific examples of applying this principle.

The first application is to use a hemodialysis device made by inserting a DNA collodione charcoal filter between two mechanical filters. The problem with this application is that the absorber is semiselective for immune components. That is, charcoal substrate non-selectively removes many of the important low molecular weight components from the treated plasma.

A second application is the removal of certain antigens using an antimicrobial fibrous coating. However, even in this application, the control coating is known to absorb a significant amount of protein non-selectively.

A third application is the semi-selective absorption of immune hemoglobin by treating the plasma with formalin and heat sterilized Staphylococcus flock. The biological action of certain strains of the Staphylococcus aureus is because a molecule called Protein A on the cell wall interacts and binds with the FC portion of mammalian IgG. However, since it interacts with the FC moiety, it does not distinguish between normal IgG and pathological IgG, and it is known that there is a possibility of serious side effects.

A fourth application is the use of a combination of filtration and cold treatment chambers to semi-selectively remove cryocytic material from plasma. However, the incidence and composition of cryoprecipitate deposits do not necessarily coincide with or indicate a variety of autoimmune or neoplastic diseases.

Another problem with the conventional method is that it does not provide a great benefit to the patient because it is not possible to selectively remove only certain pathogens to be removed without mechanical filtering. Treatment of blood and / or plasma with the methods of the present invention allows for the highly selective and economical removal of certain pathologies.

The present invention relates to a biocompatible polymer, in which a reactive biochemical is immobilized, thereby very selectively binding a component that becomes a pathogen. In the method of the present invention, a biocompatible polymer support is used, and if necessary, the spacer may or may not be attached to the biocompatible polymer support, and when attached, the size of the spacer extends from the surface of the biocompatible polymer support. Get out. One or several biochemicals are also anchored to the biocompatible polymer support or spacer by chemical bonds, which have the reactivity to bind specific pathogens or populations thereof.

The present invention also relates to the treatment of autoimmune diseases, wherein the desired pathogens are removed from the patient's blood or plasma by passing the patient's blood, plasma or other biofluid over a biocompatible polymer immobilized with reactive biochemicals. This includes the method of returning blood back to the patient. The present invention also relates to a process for the preparation of the above-described biocompatible polymers in which a reactive biochemical is immobilized, wherein the biochemical acts to selectively bind components that become pathologies. The present invention is also a method of supporting a biocompatible polymer to a mechanical support, thereby achieving good mechanical integrity.

Biocompatible Polymer Supports

The biocompatible polymer support used in the present invention, on the one hand, does not cause adverse reactions when contacted with biological fluids such as plasma or whole blood, while on the other hand it is possible to keep the reactive immobilized biochemicals extending out from the surface of the polymer support. It is a substance. Suitable materials are those that can be cast into thin films and other physical forms, yet are easy to chemically bond with the biochemicals without damaging themselves or the biochemicals. Generally, the type of material deemed suitable is called a hydrogel and can be a copolymer or an homopolymer.

Modified fibrous and fibrous derivatives, in particular cellulose acetate, are also useful as biocompatible supports of the present invention. Herein, the modified fibrous derivative is a modification of the surface of the fibrous polymer such that the biocompatibility stretched to the fibrous matrix polymer is equivalently bonded to the surface group to increase the biocompatibility. There are many suitable surfaces for the purposes of the present invention, in particular albumin being useful as a modifier. The method of attaching these groups is described later.

Homologous polymers can also be used as the biocompatible polymer support bonded to the present invention. However, the cognate polymer here includes a substance which can be judged as a slightly cross-linked cognate polymer. That is, a relatively small amount of the second component is essentially included in the production of the monomer, or is added to prevent sufficient dissolution of the homopolymer in an aqueous medium, such as blood, by sufficient crosslinking. As such an homopolymer in a drug-exchanged form is hydroxyethyl methacrylate (HEMA).

With regard to hydrogels, suitable polymers may be normal homopolymers that do not actually include other materials in the parent, or may be copolymers made from two or more monomers such as styrene and vinyl acetate. In some cases, the desired properties of the biocompatible polymeric support material can be enhanced by incorporating various monomers into the copolymer. Examples of suitable monomers for the copolymerization are hydroxyethyl methacrylate and glycidyl methacrylate.

Terpolymers belonging to the copolymer and made by polymerizing three monomers are also useful. Examples of suitable terpolymers are glycidyl methacrylate / N-vinyl pyrrolidone / hydroxyethyl methacrylate (GMA / NVP / HEMA).

In addition to the specific copolymers and homopolymers described above, copolymers suitable for the present invention (which may or may not further participate in various other monomers as required) and the homopolymers may be made by polymerizing the following monomers. Hydroxyalkyl acrylates and hydroxyalkyl methacrylates (such as hydroxyethyl acrylate, hydroxypropyl acrylate and hydroxybutyl methacrylate); Epoxy acrylate and epoxy methacrylate (such as glycidyl methacrylate); Amino alkyl acrylates and amino alkyl methacrylates; N-vinyl compounds (such as N-vinylpyrrolidone, N-vinyl carbazole, N-vinyl acetamide and N-vinyl succinimide); Aminostyrene; Polyvinyl alcohol and polyvinyl amine made from suitable polymerization precursors; Polyacrylamide and various substituted polyacrylamides; Vinyl pyridine; Vinyl sulfonate and polyvinyl sulfate; Vinylene carbonate; Vinyl acetic acid and vinyl crotonic acid; Allyl amine and allyl alcohol; Processes for preparing copolymers and / or homopolymers from vinyl glycidyl ethers and allyl glycidyl ethers, monomers described above, are well known. However, it is important in the present invention that the final copolymer and / or homopolymer should be nontoxic to animals, including humans.

The method of casting these materials into a form suitable for the present invention is of little importance. One preferred method is rotary casting, which is illustrated in Examples 2, 3 and 4.

Biochemicals

In the present invention, a biochemical is a compound which can covalently bond with a biocompatible polymer support or spacer (described below) on the one hand and bind the desired pathologic component on the other hand. In addition, the size of the biochemicals of the present invention, when covalently bonded to the polymer support, should not be too small to enter the porous substrate of the support and to form a chemical bond inside or inside the support. From this point of view, spacers can be used to ensure that the reaction site of the biochemical reaches the desired pathogen and reliably binds it. In other words, the reaction site of the biochemical is brought out from the support to be in contact with the biological fluid flowing over the support. As is apparent from the above, the action of binding the desired pathological components is in effect followed by immobilization of the biochemical to the biocompatible polymer support.

Examples of substances that can be used as biochemicals include: acetylcholine receptor protein, tissue compatible antigen, ribonucleic acid, basal capsular protein, immune hemoglobin class and subclasses thereof, spinal cord protein receptor, complement, spinal cord protein, And various specific examples of the hormones, vitamins and their receptor components include: attaching insulin to a biocompatible polymer support to remove anti-insulin antibodies associated with insulin resistance in autoimmune diseases; Attaching anti-Clg and / or Clg to the biocompatible polymer support to remove immune complexes associated with connective tissue and proliferative diseases (such as rheumatoid arthritis and cancer).

Any generally known method may be used to attach the biochemical to the biocompatible polymer support. However, biochemicals still have at least one active site, which should be used for ingredients associated with certain autoimmune diseases. In general, chemical attachment methods can be classified into the following three types. That is, 1) natural attachment, 2) chemical activation of the terminal functional group, and 3) binding reagent attachment.

The natural covalent attachment of biochemicals to the surface of the polymer support is through a chemical reactor extending from the underbody support. Reactors such as aldehydes and epoxies, e.g., extending from the polymer support, readily combine with biochemicals containing hydroxyl, amino or thiol groups. The free aldehyde groups on the polymer support also associate with hydroxyl-containing biochemicals through acetal bonds and amino-containing molecules via amide bonds. Free oxime groups are bonded to biochemicals comprising amine, hydroxyl and thio groups, respectively, via alkylamine, ether and thioether bonds. For convenience, all the attachments and couplings described above are defined as immobilization.

Chemical activation of the terminal functional groups can be carried out by chemically modifying and activating the terminal components of the polymer surface functional groups. For example, the terminal epoxy functional group is oxidized with periodic acid to form an active aldehyde group. The fixing method of the biochemical is as described above. Condensation reaction can be carried out between the free carboxyl and the amine groups through the carboxyimide activation of the carboxyl group. In other words, the carboxyl group of the polymer or bio-affinity material is carbodiimide-activated and condensed with free amine to form a stable peptide bond to fix the biochemical. The final direction of the biochemical is a factor in determining whether to use an amine-containing polymer or a carboxyl-containing polymer.

The binding reagent attachment method can be carried out by forming a covalent bond between the polymer and the biochemical using various binders. For example, free hydroxyl and / or amine containing polymers and biochemicals are covalently linked with reagents such as cyanogen bromide, diisocyanate, dialdehydes and trichloro-S-triazines.

Specifically given the preferred method of immobilizing the reactive biochemical on the biocompatible polymer substrate is generally determined by the molecular position of the biochemical and the functional groups on the polymer substrate that can be covalently bonded to the reactive bonding group of the biochemical. For example, in the case of a polymer substrate containing a terminal hydroxy functional group, it is preferable to activate by treating with an alkaline solution of cyanogen bromide (10-20% W / V). Usually the reaction mixture is kept at room temperature (20-25 ° C.) for about 30 minutes. Alkaline substances such as KOH or NaOH are added to maintain the solution in the pH range of about 10-12. The polymer is washed well with physiological saline (0.9 m%) and placed in a solution of purified biochemicals in a slightly alkaline buffer solution at 2-8 ° C. for 12-16 hours. The polymer is washed well with physiological saline to remove unbound or non-selectively bound biochemical components.

To fix biochemicals on glycidyl containing polymers, ether, thioether or alkylamine bonds are used. Epoxy-activated polymer substrates are washed with neutral buffered solution at room temperature and expanded. Purified biochemicals, dissolved borate, carbonate or phosphate buffers are placed with the glycidyl polymer substrate at 4-30 ° C. for 12-20 hours. The polymer is washed with brine, acetic acid (0.2-1.0 M) and phosphate-buffered (pH = 7.2 ± 0.2) solutions to remove excess and non-selectively bound biochemicals. Amine and carboxyl containing polymer substrates are activated by treatment with purified biochemicals dissolved in a weakly acidic (pH 4.5-6.5) buffer solution of water soluble carbodiimide. The biochemicals are covalently bound to the polymer support substrate by placing the polymer in the support, biochemicals and carbodiimide reactants at 2-8 ° C. for 12-16 hours. The polymer-biochemical combination is washed alternately with an acidic and alkaline wash solution and washed until the wash solution is free of biochemicals and carbodiimide reactants.

To determine the selective binding properties of biochemicals immobilized on the polymer, serum solutions with supplemental biomolecules were treated with active coating. The amount of biomolecules was measured radiochemically. Short exposure to biochemically modified polymer substrates resulted in significant decreases in certain biomolecules.

Spacer

In the present invention, the spacer may be defined as a molecule or compound that can attach to the surface of the biocompatible polymer support, is large enough to extend out from the surface of the support, and can fix the biochemical. The spacer keeps the active site of the biochemical outward from the support to allow for more effective contact with the biological fluid. As is apparent from the above, the action of binding the desired disease factor is actually performed after the biochemical is immobilized to the spacer, and therefore to the biocompatible polymer support.

The spacer is obtained from organic molecules in which at least two reactive functional groups are generally located at both ends of the molecule. These functional groups serve as attachment means for bonding spacers to polymer supports and biochemicals. The reactive functional groups of the spacer may be the same or different. However, these functional groups must react with the functional groups on the surface of the polymer support and also form a covalent bond with the functional groups extending from the biochemical. Any known method for carrying out this coupling reaction can be used sufficiently. For example, the above-mentioned bonding method for directly attaching a biochemical to a polymer support can be used.

Examples of suitable spacers that can be used in the present invention include those having the same reactive functional groups, such as 1,6-diaminohexane, glutaraldehyde, 1,4-cyclohexane-dicarboxylic acid, ethylenediamine tetraacetic acid, triethylene Glycols, 1,4-butanediol diglycidyl ether, methylene-p-phenyl diisocyanate and succinic anhydride and the like. Examples of spacers that do not have the same reactive functional groups include 6-aminocaprophosphoric acid, P-nitrobenzoyl chloride, 1,2-epoxy-3- (P-nitrophenoxy) -propane, aminopropyl triethoxy-silane and homocysteine Thiol acetone and the like. Polypeptides and proteins can also be used as spacers of the invention. For example, albumin of low affinity proteins has been used successfully as spacers. In addition, albumin and other natural proteins also act to increase the biocompatibility of the polymer support.

Finally, some substances act as spacers, while also acting as activators in the reaction of binding spacers and biocompatible supports. Examples of this kind are glutaraldehyde and 1,4-butanediol diglycidyl ether.

Support member

Most, but not all, of suitable biocompatible polymer supports have very low mechanical stability. Most of these materials are not high mechanical stability materials such as polypropylene thin plates, and are in fact gel or similar forms. Therefore, most of the specific forms to which the present invention is applied require a mechanically stable support member. This support member allows the use of a large surface area, enabling the rapid and medically and commercially acceptable levels of removal of immunological name components. The support member must not only be mechanically stable, but also inexpensive and sterilizable, so that the support member can be used in the device to treat the blood of the patient by the present invention. Examples of suitable materials for the support member of the present invention include filter paper, polyester fibers, polycarbonates, reticular polyurethanes, polyphenylene oxide polymers, microporous polyolefins such as polypropylene, cotton fabrics and the like.

Various methods can be used to attach a biocompatible polymer support to which a biochemical is chemically attached. For example, methods such as thin coating, horizontal casting, vacuum penetration, dip coating, crosslinking after dip coating, solution copolymerization and the like can be used. Specific examples of these methods are described in the Examples below.

cure

The therapy in the present invention refers to a therapy for autoimmunity and other diseases, the contents of which are as follows. Exposing the patient's blood or blood serum to a biocompatible polymer with immobilized reactive biochemicals removes certain pathogens and then introduces the blood back to the patient. This treatment may or may not use blood separation techniques. Therefore, this treatment can be carried out similarly to dialysis treatment, which does not require separation of whole blood or rarely physically damage normal blood components.

The method of the present invention can of course be used for the treatment of plasma. Plasma can be taken from whole blood in a number of well-known ways. Thus, for example, plasma is separated from the patient's blood by well-known methods and treated by the present invention, which is then combined with other blood components and reintroduced to the patient. In addition, for example, when obtaining plasma from a blood bank and injecting it into a patient, the plasma may first be treated by the present invention. Of course, the whole blood obtained from the blood bank can be processed by the present invention. The invention can also be used to remove pathogens from other fluids in a living body.

Because of the above and other advantages of the present invention, various types of disease states will respond to the therapy of the present invention. In general, six groups of disease states can be beneficially treated. These six disease ranges are immune component disease, drug overuse, toxin exposure, biomaterial imbalance, infection and pay-as-you-go status. Many diseases are currently being treated by plasmaperesis and cytopheresis, where the desired result is the removal of certain substances. The method of the present invention is thus applicable to diseases currently being treated by plasmaperesis and cytopheressis. Examples of immune complex diseases that can be treated are disease states including antibodies, antigens, antibody-antigens and antibody-antibody interactions, cell surface complexes, cytoplasmic complexes, and the like. Examples of drug overdose that can be treated are overdose such as iron, dioxin, aspirin, tilenol, methoxtrexate and other tricyclics.

Examples of detoxification and toxins for which the present invention is effective are lead, aluminum, mushrooms (quasi-toxin), organic phosphates, and the like. Excessive biomaterials can become a disease. Examples of such substances that can be removed using the present invention include cholesterol, uric acid, immune hemoglobin, diseased cells, uremia toxin, bilirubin, porphyrin, cortisol, and prostaglandin. Examples of infectious agents that can be treated include viral diseases such as cytomegalovirus; Prototypical diseases such as malaria, flagella, and Raysimania; Bacterial infections such as streptococci; Fungal infections such as tinea stains; Fungal plasma such as tissues such as pleural pneumonia; Rickettsian diseases such as typhoid and meningitis; Spirohetas such as mattock; And chlamydia factor in the parotid lymphatic granular-trichoma disease group.

Examples of tumors that can be treated with the present invention include lymphomas, sarcomas, cancers and leukemias. These tumors can be treated by selectively removing the cell lines, suppressors, developmental factors, and combinations thereof of the disease.

Other examples of disease states that can be treated using the present invention include the following. Postmortem staphylococcal renal nephritis, non-acute bacterial endocarditis stage 2 syphilis, pneumococcal sepsis, leprosy leprosy, abdominal tissue infection, infectious mononucleosis, typhoid, non-acute scleroencephalitis, Landry-Gilein Bar syndrome, hepatitis B infection, 4 days of fever malaria, schizophrenic infection and flagella worm infection, liver tumor, lymphoma, Hodgkins' disease, acute leukemia, multiple kidney cancer, colon cancer, bronchial cancer and Burkitz's lymphoma Tumors, arterial nodules, chronic renal glomerulonephritis, acute or non-acute thyroiditis, vinyl chloride poisoning, chronic liver disease, concomitant hypocytopenia, Burger's disease or kidney disease, rapidly developing renal glomerulonephritis and cyclic cell anemia Connective tissue diseases.

Thrombin Cytopathic Purpura, Autoimmune Anemia, Idiopathic Thrombin Cytopathic Purpura, Idiopathic Neutropenia, Cold Hemagglutinin Disease, Paroxysmal Cold Hemoglobinuria, Circulating Anticoagulant, Hemophilia, Leukemia, Lymphoma, Lupus Fetal Disease, Pernicious Anemia and Rh Disease Such as blood diseases.

Neurological diseases such as acute despinal encephalitis, multiple sclerosis, Landry's palsy, Guillain-Barzing syndrome, pseudoneuritis, and depression.

Collagen diseases such as Raynaud's disease, lupus erythema, multiple arteritis nodules, scleroderma, Sjogren's symptom group, rheumatoid arthritis, rheumatic fever, erythematous nodules.

Endocrine diseases such as Huxing's symptomatic group and diseases, thyroiditis, thyroid addiction, Addison's disease, aspermatorene.

Gastrointestinal diseases such as gastrointestinal stool, acute hepatitis, chronic active hepatitis, lupus hepatitis, cholecystosis, neoplastic colitis, local enteritis pancreatitis.

Hypercholesterolemia, renal glomerulonephritis, basal valve disease, psychogenic state-pharmaceuticals, aortic prosthetic-anemia, skin disease peeling off, Id response, dry ringworm, Behcet's syndrome, cancer, non-acute bacterial heart disease, hypertension Mixed diseases such as asthma, hereditary angina neuropathy, meningitis microosteoarthritis, Croon's disease, interstitial encephalitis and Raynaud's disease.

Diseases characterized by nuclear antigens, cytoplasmic antigens, cell surface antigens and antibodies to subclasses thereof can also be treated by the present invention. Suitable examples include the following: Native-DNA (double stranded) or antibodies against single and double strands, antibodies against SS DNA, antibodies against deoxyribonucleic proteins, antibodies against histones, antibodies against Sm, antibodies against RNPs, SC 1-1 Antibody-curable skin for, Antibody-Shogren's symptomatic group for SS-A, Sykka complex, Antibody-Rheumatoid Arthritis for RAP, Sjogren's symptom group, Antibody-Multiple Dilapidosis-Skin Dermal Shrinkage Syndrome, and antibody-histosclerosis against nucleus phosphorus, Sjogren's group of signs.

In addition, antibodies related to certain autoimmune diseases include, for example, antibody-chronic hepatitis to hairless muscle, antibody-depression to acetylcholine receptors, antibody to cataracts at the dermis-epidermal junction, mucopolysaccharides Antibody-Solaris against protein complex or internal cell cement material, Antibody-Rheumatoid arthritis against immunoglobulin, Antibody-nephrocyte nephritis against renal glomerular base film, Goodpasture symptomatic group, Sudden stage 1 hemasidero Cis, antibody-autoimmune anemia to erythrosite, antibody-hacimodian disease to thyroid, antibody-malignant phenotype to essence factors, antibody-acute thrombin cytoplasmic purpura to platelets, macroimmunization, antibodies to mitochondria- Ciliary cirrhosis, antibody-Sjogren's symptom group for salivary gland cells, antibody-abrupt adrenal atrophy for adrenal gland, thyroid particulates The antibody-Graves disease when antibodies for thyroid hyeolguso - Addison's disease, antibodies to the Isle Inlet cells diabetes.

Hyperprotein diseases such as multiple myelitis. Solid cellulosis, hypocytopenia and mild chain disease.

Hyperlipidemia, such as stage 1 cystic cirrhosis and hypercholesterolemia.

Endocrine diseases such as Graves' disease and diabetes.

Symptomatic immunization, eg, neonatal blood disease and kidney transplant rejection.

Suitable for treatment with the present invention are also post-transfusion purpura and autoantibodies, for example: Goodpasture's symptomatic group, depression, malignant swelling, blood disease, sudden (autoimmune) thrombin cytoplasmic purpura, autoimmune anemia, control factor for factor V111, multiple myopathy / Guillain-Barring syndrome.

Immune complex diseases can also be treated, for example: Tissue lupus erythema, multiple arteritis nodules, cutaneous vasculitis, rheumatoid arthritis, renal glomerulonephritis and cutaneous pupillary shrinkage.

While it is not possible to turn to any one theory, looking at the progression of autoimmune pathologies, the sequence usually begins with the challenge of free antigens, antibodies develop, and after they form a complex, antibody surpluses and supplements form complexes. Coagulated. Therefore, preliminary diagnostic procedures to determine the dominant form of autoimmune factors are needed to properly select biocompatible polymer structures to increase efficacy. Next, the treatment of rheumatic diseases will be described by way of example. Rheumatic diseases can usually be characterized by the following processes: RF complex coagulation with a) free antigen (abnormal Ig) (rheumatic state), b) free RF antibody generation and RF complex formation, and c) activation of antibody excess and supplement. Thus, treatment at an early stage of development of rheumatoid disease can be determined by detection of abnormal immune hemoglobin, a monolithic rheumatoid factor (m-RF) antibody. Treatment at this stage can be carried out very effectively by removing the infiltrating antigen by the m-RF activating biocompatible polymer to prevent the development of endogenous RF (e-RF) antibodies. Diagnostic detection of e-RF indicates the use of biocompatible polymers having both m-RF and aggregate gamma hemoglobin active biochemicals.

Alternatively, two biocompatible polymers each having one type of active biochemical may be used in sequence. In either case, by combining m-RF and aggregate gamma hemoglobin, both penetrating antigen and antibody molecules are absorbed to prevent disease progression. Where significant levels of RF antigen-antibody complexes are detected, biocompatible polymers comprising Clq and / or collagen factor molecules are indicated. Finally, if the disease has progressed to the fixation stage of the immune complex, an effective biocompatible polymer will comprise one or more anti-supplement antibodies (eg anti-Clq, anti-C ₃ or anti-C ₄ ). In addition, when more than one biochemical is desired, they may be immobilized on one biocompatible support, or each may be immobilized on a separate support to connect in turn with respect to the flow of blood or plasma.

As pointed out above, effective use of the present invention requires well-defined driving forces and stages of immune response to disease management.

Today plasmapheressis and sight peresis treat diseases by removing toxins or cells from the blood. Diseases currently being treated by the removal of certain substances by plasmaperesis and / or cytoperesis can be better treated with the products and methods of the invention. Specifically, the treatment method of the present invention for whole blood can be explained as follows: a) by installing a vascular passageway, b) allowing blood to flow at about 30 ml / min to 200 ml / min, and c) administering an anticoagulant into the blood. D) neutralizing the anticoagulant effect on the treated blood by d) installing a pumping device, e) passing the blood in accordance with the present invention, and f) administering additional drugs according to the anticoagulant used, g) reimbursing the patient for the blood Put it in.

The time range required for the above treatment is about 2 to 4 hours. Of course, depending on the situation, you can shorten or extend the time. The treatment of the present invention for plasma can be described as follows. a) establish a vascular pathway, b) allow blood to flow from about 30 ml / min to 200 ml / min, c) administer anticoagulant to the blood, d) install a pump device, and e) plasma-forming blood component separation device. F) passing the plasma according to the invention, g) filtering through a 0.2 micron filter to remove microembolites, bacteria and / or fungi, h) recombining the treated plasma with other blood components, i) Additional drug administration, depending on the anticoagulant used, neutralizes the anticoagulant effect on the treated blood, j) reintroduces the treated blood to the patient.

Methods of installing vascular passageways are well known in the medical community. For example, a cannula with a large hole in the stomach can be used intravenously or as an artery. Examples of suitable veins and arteries are anterior veins, subclavian veins, and arm or radial arteries. Arterial venous fistulas (AV shunts) may also be used. In this case, the heart is the pump device. When no AV fistula is used, a preferred pump device is a roller type communication pump that can supply about 30 ml / min to 200 ml / min.

Examples of anticoagulants suitable for the method of the present invention include a mixture of acid citrate glucose (approximately 1 ml per 8 ml of total blood), heparin, heparin / acid citrate grapes (i.e. 1250 IU of heparin in 125 ml of acid citrate glucose). And prostaglandins. When using anticoagulants such as heparin and prostaglandins, it is common to administer neutralizing medications before reinjecting the therapeutic blood or plasma into the patient. When treating plasma, conventional methods can be used to remove other components of the blood. Examples of methods for separating plasma from other blood components include plasma peresis, cell centrifugation, and cell precipitation in plasma bags. If necessary, both continuous separation and single (batch) separation can be used, and this separation is not related to the present invention.

Finally, the form of the present invention is largely not important. For example, the biocompatible support of the present invention may be used in the form of plate, concave fibrous, cylindrical fibrous, reticular, cylindrical or rectangular passage, necklace shape, and the like, and the combination thereof. It may be beneficial to use a fluidized bed for some diesel.

Example 1

This example describes a method for predominantly biocompatible polymer supports and chemically attaching biochemicals to direct polymer supports. This embodiment also illustrates the use of a device without mechanical support.

Absorption of anti-insulin antibodies using insulin activated poly hydroxyethyl methacrylate (P-HEMA) coatings.

A. Polymer Structure

15.0 g of 2-hydroxyethyl methacrylate, 15.0 g of ethylene glycol, 0.08 g of sodium bisulfite, and 0.036 g of ammonium persulfate were combined to form a monomer solution. The solution was stirred at rt for 15 min. Cut a polyethylene spacer (10 mils thick) in the center of the glass plate (5 "long x 5" wide x 3/8 thick) to form a gasket frame to make a 4 "x 4" glass window, and add about 5 ml of the solution Poured. A second glass plate was placed on the gasket and solution, clamped and the whole device was left at 60 ° C. overnight. The cramps were removed, the glass plate was slightly lifted up and transferred to a non-ionized bath for at least 24 hours. The swollen polymer thin film from the glass plate was carefully removed and hydrated by washing with fresh water that was not ionized for at least 3 days. (500ml / day)

B. Polymer Activation

The polymer valve was cut to make a thin film disc (5 mm in diameter) for activation and analysis. 10 ml of 0.2M Na ₂ CO ₃ (pH = 11.1) was dissolved at 1.69 g of fine powder BrCN crystals under continuous stirring at 4 ° C. to prepare a 10-20 gm% brominated cyanogen solution. 5N NaOH was added dropwise until the crystals were dissolved to maintain the pH of the solution at 11 or higher and to stabilize the pH. Four thin film discs were placed in small portions, washed with about 5 ml of 0.1N HCl, and then placed in a cyanobromide solution for 15 minutes. The discs were washed at least twice with 5 ml each of 0.1N HCl and placed overnight in 5.0 ml of U-100 regular ILETIN insulin solution (adjusted pH to 8.7 by addition of 1N NaOH). The thin film disc was washed with 5 ml of 0.5 M NaCl, 0.1 M Na ₂ CO ₃ solution, and 5 ml each of phosphate (0.05 M) buffered saline (0.9 gm%) solution (pH = 7.4).

C. Evaluation of Thin Film Uptake of Anti-Insulin Antibodies

A series of dilutions (from 980 pg to 15 pg) of P-HEMA thin film discs with 560 pg (picograms) of I ¹²⁵ labeled porcine insulin and no-labeled porcine insulin, ie 280 pg guinea pig anti-swine insulin antibody, was used. The double antibody competitive radioactive immunoassay was carried out by placing in 0.5 ml of phosphate (0.05 M) buffer (pH = 7.49) saline (0.9 gm%) containing serum albumin at room temperature for 2 hours. P-HEMA discs were removed from each test solution. 0.1 ml of goat anti-guinig gamma hemoglobin was added to each test tub. The test solutions were mixed and left for another 2 hours at room temperature. 0.1 ml of cold (2-4 ° C.) phosphate buffered saline (pH = 7.4) was added to each tub. Each test solution was mixed and centrifuged at 7500 G for 15 minutes at 4 ° C., then the suspension was poured into 20 ml scintillation bottles. 5.0 ml of aquasol age scintillation fluid was added to make the suspension gel and measured for 4.0 minutes in an Isocop 300 counter. The insulin treated thin film discs absorbed 111 pg of anti-insulin antibody from the solution, ie 690 pg of anti-insulin antibody per cm 3 of surface area.

Example 2

This example illustrates how to make an unsupported biocompatible thin film. This example also shows how to use 6-aminocaprophosphoric acid (6-carbon chain) as a spacer, how insulin is attached to a biocompatible polymer support to remove insulin antibodies, and also insulin activated polyhydroxyethyl methacryl A P-HEMA thin film is used to explain how to absorb anti-insulin antibodies.

A. Polymer Casting

15.0 g of 2-hydroxyethyl methacrylate, 15.0 g of ethylene glycol, 0.08 g of sodium bisulfite, and 0.036 g of ammonium persulfate were combined to form a monomer solution. The solution was stirred at rt for 15 min. Cut a polyethylene spacer (10 mils thick) in the center of the glass plate (5 "long x 5" wide x 3/8 ") to form a gasket to make a 4" x 4 "window, and about 5 ml of solution Place the second glass plate on top of the gasket and solution, tighten it with a bracket, and let the whole device stand overnight at 60 ° C. Remove the bracket, remove the glass plate a little and move it to the non-ionized bath at least 24 The swollen polymer thin film was carefully removed from the glass plate and washed-hydrated for at least 3 days (500 ml / day) with fresh non-ionized water.

B. Polymer Activation

A thin film disc was made as in Example 1. 10 ml of 0.2 M Na ₂ CO ₃ (pH = 11.1) was dissolved in 1.69 g of finely divided BrCN crystals with continued stirring at 4 ° C. to accommodate 10-20 gm% cyanogen bromide solution. 5N NaOH was added dropwise until the crystals were dissolved to maintain the pH of the solution at 11 or higher and to stabilize the pH. Four thin film discs were placed in small portions, washed with about 5 ml of 0.1 N HCl, and placed in cyanogen bromide solution for 15 minutes. The discs were washed at least twice more with 5 ml each of 0.1N HCl, overnight in 10 ml of 10gm% solution (W / V) in which 6-aminocaproic acid was dissolved in 0.1 M Na ₂ CO ₃ , 0.5 M NaCl buffer pH 8.6. I put it in. The thin film disc was washed with 5 ml of 0.1 M Na ₂ CO ₃ , 0.5 M NaCl buffer and 3 times of 5 ml each of phosphate (0.05 M) buffer (pH = 7.4) saline (0.9 gm%) solution. The thin film disc was removed from the washing solution, and cyclohexyl-3- (2-morpholinolethyl) capbodiimide was dissolved in 0.1M 2 (N-motopolyno) ethanesulfonic acid (MES) buffer (pH = 6.0). 10 ml of 10 gm% solution (W / V) was activated for 30 minutes at room temperature, and then each disc was washed in 5 ml of cold (4 ° C) phosphate buffered saline solution. The same thin film discs were placed overnight at 4 ° C. in 5.0m V-100 normal ILETIN insulin injection solution or porcine insulin normal ILETIN solution. The thin film disc was removed from the protein solution and washed three times in 5 ml of phosphate buffered saline solution.

C. Evaluation of Thin Film Absorption of Anti-Insulin Antibodies

Contain a series of dilutions (980pg to 15pg) of 560 pg I ¹²⁵ labeled porcine insulin and unmarked porcine insulin, ie 280 pg guinea pig anti-pig insulin antibody, containing 1 gm% bovine serum albumin. In a 0.5 ml phosphate (0.05 M) buffer (pH = 7.4) saline (0.9 gm%), a double antibody competition-radioactive immunoassay was performed by placing at room temperature for 2 hours. The P-HEMA disc was removed from each test solution. 0.1 ml of goat anti-guinea pig gamma hemoglobin was added to each test tub. The test solutions were mixed and held for another 2 hours at room temperature. 1.0 ml of cold (2-4 ° C.) phosphate buffered saline (pH = 7.4) was added to each tub. Each test solution was mixed and centrifuged at 7500 G for 15 minutes at 4 ° C., then the suspension was poured into 200 ml scintillation bottle. 5.0 ml of aquasol liquid scintillation fluid was added to make a gel and measured for 4.0 minutes in an Isocap 300 counter. Insulin-treated thin film discs absorbed 271 pg of anti-insulin antibody from the solution, ie 690 pg of anti-insulin antibody per cm 2 of surface area.

Example 3

This embodiment describes a method of casting a biocompatible polymer support by rotary casting, and includes both a case of using and not using a mechanical support member. This example also describes a second method for chemically bonding biochemicals to biocompatible polymer supports.

Absorption of Anti-Human Hemoglobin G (IgG) Antibody (Rheumatoid Factor) Using Immune Hemoglobin-Activated Poly-hydroxyethyl Methacrylate-and-Glycidyl Methacrylate (P-HEGL) Thin Films

A. Polymer Casting

The following example illustrates a method for producing a polymer thin film supported and unsupported polymer thin film by rotary casting.

a. Rotary casting machine.

The rotary casting machine consists of a closed aluminum drum with a 1/4 inch wall. The drum's internal dimensions are 4 inches in diameter and 5 inches in length. The drum is connected to the motor to rotate, and the rotational speed (rpm) of the drum is measured with a strobe phototameter. The hot air gun heats the rotating drum, and the thermocouple measures the internal temperature of the drum and the temperature of the air flowing over the drum. The drum is flushed with nitrogen before and during the polymerization.

b. Preparation of Supported Valves.

Hard-loading preliminary paper of Wattram class 50 was used as a supporting member to give mechanical force against rotational casting. The filter paper was cut into rectangular pieces 4-15 / 16 × 12-7 / 16 inches) and then soaked for 30 minutes at room temperature in ethylene glycol (EG). When the excess glycol was removed, the filter paper contained 2-4 g of EG. The filter paper thus treated was rolled into a cylindrical shape and placed in a rotary casting drum. The outer end of the filter paper was pressed against the wall of the drum, so that all the air between the wall and the filter paper escaped. Although not required, it is desirable that both ends of the filter paper come into contact with each other when it is in place. They may overlap each other. The filter paper member was checked for air bubbles. If there were bubbles, they were removed with a rubber rod.

In polymerizations that produce highly adhesive polymers, the rotary casting cylinder may first be coated with silicon release paper. Next, Whatman filter paper treated as described above may be placed in contact with the discharged paper. However, care must be taken not to trap air.

C. Polymerization Composition

The following is the polymerization composition currently used. In each case the animal was stirred with the reactive monomer for 30 minutes at room temperature, ie until the animal dissolved.

GMA-HEMA (50/50) Copolymer

6.25 g 2-hydroxyethyl methoxylate (HEMA)

6.25 g glycidyl methacrylate (GMA)

12.5 g ethylene glycol (EG)

0.02 g 2,2'-azobis (2-amidinopropane)

Hydrochloride (ABAP)

GMA-NVP-HEMA (50/40/10) Copolymer

6.25g GMA

1.25g NVP

5.00g HEMA

12.5g EG ^*

0.02g ABAP

Ethylene glycol weights include EG2-4g on Whatman filter paper.

d. Rotary casting method.

The drum was installed in a rotary casting machine while the animal was dissolved in the monomers. The drum was spun at 1400 rpm at room temperature, rinsed with nitrogen for 15 minutes, and then injected into the drum with an animal-monomer solution (25.0 ml) with a hypodermic syringe. The tip of the syringe is made of flexible teflon. Rinse again with nitrogen and increase the drum speed to 2,900 rpm.

The drum was heated at 70-75 ° C. for 90 minutes by operating a blower (approximately 35 ft ³ / min) and a heater attached to the heat blow gun. The heat was then cut off and the blower was left to operate, cooling the drums to bring the internal temperature down to about 30 ° C. The cooled drum was removed from the rotary casting machine and filled with non-ionized water. After soaking for 1 hour, the casting was processed from the drum.

B. Polymer Activation

A thin film disc was made as in Example 1. Fourteen individual discs were placed in 1.0 ml of 1.0 M hexanediamine solution at 4 ° C. for 72 hours. The disc was removed from the hexanediamine solution and washed three times with 2 ml of phosphate buffered saline solution. A 4.0 gm% HGG solution was made by dissolving 4.0 g of human gamma hemagglutinin (HGG) while gently stirring 100 ml of 0.1 M MES buffer (pH = 6.0) at room temperature. After complete dissolution of the protein, 1.0 ml of protein solution was continuously transferred to 9.0 ml of MES solution, so that the concentration of the protein in the buffer was 4 mg / ml, 400 µg / ml, 40 µg / ml and 4 mg / ml. A diluted solution of was obtained. Two polymer discs were placed in 0.5 ml of protein solution and 0.5 ml of 0.25 M 1-ethyl-3- (3-dimethyl-aminopropyl) carbodiimide solution (solution made from MES buffer) for 72 hours at 4 ° C. Put it. Each thin film disc was removed from the protein solution and washed three times with 2 ml of cold (4 ° C.) phosphate buffered saline.

C. Evaluation of Thin Film Absorption of Anti-IgG Antibody from Physiological Solution

Radioimmunoassays were performed by placing individual thin films with 10 mg I ²⁵ goat anti-human IgG in 1.0 ml PBS containing 1.0 mg% human serum albumin for 2 hours at room temperature. The radiotracer solution was removed and each thin film was washed three times with 2.0 ml of PBS solution. The thin film was placed in the last wash solution overnight at 4 ° C. Each thin film was removed from the wash solution and measured for 1 minute each with an Innotron Hydra gamma counter. The conversion per minute (DPM) was divided by the number of times per minute divided by the detector efficiency. The average DPM was divided by the radiotracer specific gravity to approximate the amount of antibody absorbed. The following results were obtained.

Picogram of radiotracer per 1 cm 2 thin film.

** Basic

Example 4

This example shows the use of amino caprophosphate as a spacer for gamma-hemocytes.

Absorption of Anti-Human Immune Thyro G (IgG) Antibodies Using Immune Hemoglobin Activated Poly-hydroxyethyl Methacrylate- and -Glycidyl Methacrylate (P-HEGL) Thin Films:

A. Polymer Casting

Rotary cast P-HEGL thin films were made as in Example 2.

B. Polymer Induction and Activation

A thin film disc was prepared and treated in the same manner as in Example 2 except that 1.0M 6-amino caprophosphate was used instead of hexanediamine as the inducer and spacer.

C. Evaluation of uptake of thin films of anti-IgG antibodies from physiological solutions.

Radioimmunoassay was carried out in the same manner as in Example 3, and the following results were obtained.

Picogram of radiotracer per 1 cm 2 thin film.

Example 5

This example shows the use of albumin (67,000 MW) as a spacer for folate. Folate is used to remove hydrochloric acid binding proteins.

Absorption of hydrochloric acid binding protein (FABP) by folate-albumin activated poly hydroxyethyl methacrylate (P-HEMA) thin film:

A. Polymer Casting

The filter paper support P-HEMA polymer thin film was rotatably cast in the same manner as in Example 2 using the following polymer composition.

15.0g 2-hydroxyethyl methacrylate

15.0 g ethylene glycol

0.08g Sodium Metabisulfite

0.036 g ammonium persulfate

B. Polymer Induction and Activation

The bovine hydrochloride complex was made by carbodiimite condensation of a folate carboxyl group with an albumin end amine group. To do this, 200 mg of hydrochloric acid was dissolved in 8 ml of 0.1N NaOH, and 400 mg of 1-cyclohexyl-3 (2-morpholinoethyl) -carbodiimidemetho-P-toluene sulfonate was dissolved in 2.0 ml of 0.1 M MES buffer. (PH = 6.0), and 1.0 g bovine serum albumin (BSA) was also dissolved in 4000 ml of 0.1 M MES buffer. These solutions were mixed and left at 4 ° C. for 72 hours. 20 ml of the mixture was treated with 20 ml of BSA (2.5 gm%)-charcoal (1.25 gm%) suspension at 4 ° C. for 30 minutes to remove unreacted plates and carbodiimide from the solution. The suspension was poured off by centrifugation at 3400G for 15 minutes at 4 ° C. and filtered through 0.22 micron filter paper.

A thin film disc was prepared in the same manner as in Example 1 and treated with a cyanogen bromide solution. The plates were washed with cold saline solution, and then eight disks were placed in 20 ml of physiological saline, 160 gm% BSA or folate-albumin complex solution made as above and placed at 4 ° C. for 72 hours. Eight thin film baths were removed from the solution to wipe off the water, and washed in at least 4 hours at 4 ℃ in 20ml brine solution. The same thin bath was treated with 8 ml of 1% gluter aldehyde solution for 1 minute and washed overnight with 20 ml of phosphate buffered saline buffer.

C. Evaluation of Thin Film Absorption of Hydrochloric Acid Binding Protein (FABP) from Physiological Solution

370 pg of ³ H-pteroglutamic acid (PGA) and non-radioactive PGA standard dilution (48 pg to 348 pg), or 234 pg of P-HEMA thin film disc with FABP binding, 20 ul folate-free normal human serum Competition protein binding radioactivity was carried out by putting in 1.0 ml of 0.05 M phosphate buffer (pH = 7.6) containing 5 mg of sodium ascorbate. The radiometric tubings were combined and left for 30 minutes at room temperature and 10 minutes at 4 ° C. Individual thin film discs were removed from the test solution and 0.5 ml of cold (4 ° C.) BSA (2.5 gm%)-charcoal (1.25 gm%) suspension was added to each tub. All test solutions were placed at 4 ° C. for 10 minutes and centrifuged at 2000 ° C. for 15 minutes at 4 ° C. The suspension was poured into 20 ml scintillation bottles. 12.0 ml of liquid scintillation fluid was added to each bottle. Samples were taken and measured with an isocap 300 counter. The following results were obtained.

The embodiments described above represent specific forms of the present invention, but the present invention is not limited thereto. Those skilled in the art can make various changes and improvements without departing from the basic principles of the present invention.

Claims (19)

A biocompatible polymer comprising a biocompatible polymer support and one or several biochemicals that are chemically bound to and fixed to the support, wherein the biochemical is responsive to bind certain pathologies . It consists of a biocompatible polymer support, a spacer covalently bonded to the support, and one or several biochemicals immobilized by a chemical bond to the spacer, wherein the biochemical is capable of identifying a specific pathology or a specific group of pathogens. A bioselective polymer having reactivity for binding. The bioselective polymer of claim 1 or 2, wherein the biocompatible polymer support is a hydrogel. 3. The bioselective polymer of claim 1 or 2, wherein the biocompatible polymer support is a slightly crosslinked homopolymer hydroxy ethyl methacrylate. 3. The bioselective polymer of claim 1 or 2, wherein the biocompatible polymer support is selected from the group consisting of polymerized glycidyl acrylate and polymerized glycidyl methacrylate. The biocompatible polymer support of claim 1 or 2, wherein the biocompatible polymer support is selected from the group consisting of copolymerized N-vinyl pyrrolidone and glycidyl methacrylate, wherein the copolymer is hydroxy alkylacrylate, Bioselective polymers comprising monomers selected from the group consisting of hydroxy alkylmethacrylates, acrylamides, substituted acrylamides, vinyl glycidyl ethers, allyl glycidyl ethers, N-vinylamides, vinyl acetate and mixtures thereof . The biocompatible polymer according to claim 1 or 2, wherein the polymer support is a tertiary polymer of glycidyl methacrylate, N-vinyl pyrrolidone and hydroxyethyl methacrylate. The biocompatible polymer according to claim 1 or 2, wherein the polymer support is modified cellulose acetate. The bioselective polymer according to claim 1 or 2, wherein the polymer support is fixed to a mechanically stable support member. 10. The method according to claim 9, wherein the support member stabilized by the machine jug is purified by cellulose acetate, polyester fiber, microporous polyolefin, cotton fabric, polystyrene, polycarbonate, polyphenylene oxide, reticulated polyurethane, and combinations thereof. Bioselective polymer selected from the group consisting of. The method according to claim 1 or 2, wherein the biochemical is acetylcholine receptor protein, tissue compatible antigen, ribonucleic acid, basal thin film protein, immune hemoglobin and subclass, myeloma protein receptor, complement, myelin protein, hormone and its A bioselective polymer selected from the group consisting of receptor components, vitamins and their receptor components and the like. The bioselective polymer according to claim 1 or 2, wherein the biochemical is insulin used to remove anti-insulin antibodies associated with insulin resistance, which is an autoimmune disease. The bioselective polymer according to claim 1 or 2, wherein the biochemical is purified gamma hemoglobin for removing immune components related to connective tissue and proliferative diseases such as rheumatoid arthritis and cancer. The method of claim 2, wherein the spacer is 1,6-diaminohexane, glutaraldehyde, 1,4-cyclohexane-dicarboxylic acid, ethylene diamine tetraacetic acid, triethylene glycol, 1,4-butanediol diglyci Diether, methylene-p-phenyl diisocyanate, 6-amino caproic acid, p-nitro benzoyl chloride, 1,2-epoxy-3- (p-nitrophenoxy) propane, amino propyltriethoxy-silane, succinic acid A bioselective polymer selected from the group consisting of anhydrides, homoacid thiolactones, albumin and the like. By contacting a biofluid of a patient with a bioselective polymer consisting of a biocompatible polymer support and a reactive biochemical immobilized to the support, the biochemical can be used to determine a specific pathology or a specific population of pathologies associated with the disease state of the patient. Binding, and re-injecting the biofluid into the patient. By contacting a biofluid of a patient with a bioselective polymer consisting of a biocompatible polymer support, a spacer attached to the support, and a biochemical fixed to the spacer, the biochemical can be applied to a specific condition associated with the disease state of the patient. Binding a pathology agent or a particular group of pathogens and re-injecting the biofluid into the patient. Prior to administering to the patient a biofluid to be administered to the patient, specific pathogens are removed from the biofluid by contacting the biocompatible polymer support and the bioselective polymer immobilized thereon. And then introducing the biofluid to the patient. Before administering to the patient a biofluid to be administered to the patient, the biofluid is contacted with a biocompatible polymer comprising a biocompatible polymer support, a spacer attached to the support, and a biochemical fixed to the spacer. Removing certain pathologies from the liquid, and then introducing the biological fluid into the patient. 19. The method according to any one of claims 15, 16, 17 or 18, wherein two or more bioselective polymers are used in turn, and each of these polymers is immobilized with the same or different reactive biochemicals or populations thereof. Removing said specific pathogen.