WO2018144505A1 - Device and method for treatment of inflammation by extracorporeal peripheral blood filtration for specific removal of inflammation factors - Google Patents

Abstract

Methods and devices for treating inflammation by specifically removing inflammation factors by extracorporeal peripheral blood filtration are provided. The extracorporeal peripheral blood filtration devices generally include blood inlet and outlet ports, and a filtration matrix configured to specifically remove inflammation regulating cytokines from blood flowing through the device. The methods of treatment generally include flowing blood through an extracorporeal peripheral blood filtration device to remove inflammation regulating cytokines from the blood.

Description

DEVICE AND METHOD FOR TREATMENT OF INFLAMMATION BY EXTRACORPOREAL PERIPHERAL BLOOD FILTRATION FOR SPECIFIC REMOVAL OF INFLAMMATION FACTORS

CROSS REFERENCES TO RELATED APPLICATIONS

[0001] This application claims priority benefit of U.S. Provisional Application No.

62/452,551, filed January 31, 2017. The entire contents of which is incorporated by reference herewith.

TECHNICAL FIELD

[0002] The present invention relates to novel devices for treating inflammation associated with hypercytokinemia and commonly referred to as cytokine storm. The device is used in conjunction with dialysis instruments in patients at risk of or suffering from such inflammation via beads and/or strips or high surface area matrix that is coated with a high affinity binding antigen for cytokines or the interleukin family of molecules specifically involved in inflammation due to trauma or infection by a foreign agent such as Influenza A. Also described are therapeutic methods including filtration of peripheral blood from patients at risk of or suffering from hypercytokinemia utilizing high affinity capture beads, strips or high surface area matrix bound with inflammatory binding antigens.

BACKGROUND

[0003] Inflammation is part of the body's immune response aiming to remove harmful stimuli, including damaged cells, irritants, or pathogens. However, inflammation may become unregulated in severe infection or trauma, and in order to begin the healing process and return the injured tissue to its normal structure the inflammatory response and regulation becomes critical to achieve as a continued inflammatory response or cytokine storm could lead to tissue damage or death.

[0004] Inflammation typically begins at a local site and spreads throughout the body via systemic circulation, triggering an inflammatory response. Depending on the originating site and the stimuli causing the inflammation, the inflammatory responses vary. [0005] Once triggered, the inflammatory response is typically highly regulated by the interaction of both pro-inflammatory or anti-inflammatory stimulators and mediators. Cytokines are one class of inflammation regulatory proteins.

[0006] Generally, cytokines are a diverse class of proteins secreted for the purpose of intercellular signaling and communication. Cytokines, through receptor binding can elicit a variety of responses, depending on the cytokine itself, concentration, presence or absence of other cytokines, and the target cell. During an inflammation response, upon exposure to harmful stimuli, as part of the body's immune response cytokines are secreted for the purpose of signaling and activating immune cells such as T-cells and macrophages. Additionally, cytokines can stimulate a variety of cells to produce more cytokines.

[0007] Normally, the body keeps this self-synthesis process in check. However, in some instances, the reaction becomes uncontrolled, and too many immune cells are activated in a single place. This phenomenon has come to be known as a "hypercytokinemia," "cytokine storm," or "cytokine cascade event." These cytokine storms have potential to do significant damage to body tissues and organs. As a simple, yet illustrative, example; as a consequence of a cytokine storm fluids and immune cells such as macrophages may accumulate in the lungs, causing the airways to be blocked, potentially damaging the lung tissue or even death if severe enough.

[0008] During a cytokine storm event, the regulation of cytokine production may become disordered, resulting in a systemic over expression of normal, healthy and vigorous inflammatory mediators such as cytokines. Normal local expression of inflammation mediators, whether pro-inflammatory or anti-inflammatory, in tissue can be protective and restorative, whereas systemic overexpression typically is not. Higher than normal concentrations of certain cytokines can cause disease, tissue damage, and other deleterious health effects, some potentially lethal.

[0009] Cytokine storms can be triggered by a number of medications, infection by a variety of microorganisms, including bacteria, viruses, fungi, and parasites, and infectious, and non-infectious diseases. Cytokine storms can be triggered, for example, by severe acute respiratory syndrome ("SARS"), sepsis, Ebola, or influenza. Generally, any condition or event that would trigger an inflammatory response such as tissue injury (e.g. surgery, accidents), shock (e.g. hemorrhagic, cardiogenic, septic), or ischemia can ultimately trigger a cytokine storm event.

[0010] The ability to manipulate and correct a disordered inflammation response would be beneficial to the medical profession. However, a systemic manipulation of the inflammation response can be risky given its inherent protective and restorative functions.

[0011] Current methods to regulate and correct a disordered inflammation response do not target specific cytokines, but rather, they indiscriminately remove all cytokines. Current methods of removing cytokines utilizing size exclusion filtration as well as non-specific surface adsorption.

[0012] Generally, size exclusion filtration will indiscriminately remove the majority of cytokines because despite their diverse functions most cytokines are within a narrow -40 kilodaltons molecular weight range.

[0013] Further, adsorption of low molecular weight proteins to hemodialysis membranes also indiscriminately removes cytokines as the predominant mechanism by which these membranes remove cytokines in clinical practice is nonspecific surface adsorption.

[0014] From this perspective, therapy aimed at reducing the inflammatory response by removal of a broad spectrum of inflammatory regulating cytokines may not restore normal immune responsiveness and inhibit protective and restorative functions of a normal inflammatory response. Further, treating all disordered inflammatory responses with a "one-size-fits-all" approach will be more likely to remove both beneficial as well as detrimental cytokines. Instead, a more desirable inflammatory response modulating strategy is to use a customizable method to selectively remove pro-inflammatory and/or anti-inflammatory stimulators or mediators causing the disordered inflammatory response.

[0015] While currently available methods may have some success treating certain inflammatory conditions, there is a need in the art for a simple method that has the ability to only remove a therapeutically effective amount of only the desired cytokines.

[0016] The current invention utilizes high affinity binding to remove specific cytokines, thus providing a higher degree of control as compared to methods utilizing size exclusion or non-specific binding to remove cytokines. SUMMARY OF THE INVENTION

[0017] It is the object of this invention to overcome the challenges in treating disordered inflammatory responses. Accordingly, devices and methods for modulating an inflammatory response are disclosed herein, which are customizable to a specific patient and individual inflammatory responses.

[0018] The present invention provides devices and methods for removing or reducing levels of specified cytokines in peripheral blood in situations where abnormally high levels of one or more cytokines, or unregulated or excessive interaction among such cytokines occur, or during events that do induce or have the potential for inducing abnormal levels of one or more cytokines. The devices and methods serve to prevent, control, reduce, modulate, or alleviate the severity of the disordered inflammatory response.

[0019] Devices and methods are disclosed that remove one or more cytokines involved in the inflammatory response from peripheral blood of the patient and are useful for treating patients with a disordered inflammatory response related to either acute or chronic inflammation. Diseases or conditions related to either acute or chronic inflammation include, but not limited to, sepsis, septic shock, systemic inflammatory response syndrome ("SIRS"), severe acute respiratory syndrome ("SARS"), viral infections, bacterial infections, tissue injury, ischemia, allergic reaction, blunt force trauma and chemical or biological agent exposure.

[0020] In embodiments, the invention provides an extracorporeal peripheral blood filtration device, comprising a housing configured to receive peripheral blood, as part of dialysis or other peripheral blood filtration methods, through a peripheral blood introducing port, contacting the peripheral blood with a filtration matrix, and a peripheral blood outflow channel, wherein: the filtration matrix can include, but is not limited to, at least one capture antibody, or a functional portion thereof, immobilized on a plurality of beads, strips, or high surface area configuration or matrix.

[0021] In one aspect, the at least one capture antibody has a high affinity for one or more cytokines involved with inflammatory response such as TNF-a, IL-la, IL-Ιβ, and IL-2- IL18 but not limited to these direct response molecules. The at least one capture antibody can effectively bind and remove one or more of TNF-a, IL-la, IL-Ιβ, IL-2- IL18, or any other specified cytokine or interleukin involved in inflammation.

[0022] In embodiments, the extracorporeal peripheral blood filtration device is fitted with connective adaptors so the device can be connected in-line to existing dialysis or other peripheral blood filtration systems. The connective adaptors can include, but are not limited to, conventional mating luer fittings, enabling incorporation into existing dialysis cartridges or tubing.

[0023] In other embodiments, an extracorporeal peripheral blood filtration device is provided that includes a housing configured to receive peripheral blood, as part of a separate and distinct peripheral blood filtration system, through a blood introducing port, contacting peripheral blood with a filtration matrix, and a peripheral blood outflow channel, wherein: the filtration matrix can include, but is not limited to, at least one capture antibody immobilized on a plurality of beads, on strips, or on other high surface area matrices.

[0024] In other embodiments, an extracorporeal peripheral blood filtration device is provided that includes two or more housings sequentially connected and configured to receive peripheral blood through a blood introducing port, contacting the peripheral blood with a filtration matrix of one housing, followed by contacting the peripheral blood with a filtration matrix of the additional housings in sequence, and a peripheral blood outflow channel, wherein: the filtration matrix of each housing can include, but is not limited to, at least one capture antibody immobilized on a plurality of beads, on strips, or on other high surface area matrices.

[0025] In one aspect, the at least one capture antibody has a high affinity for one or more cytokines involved an inflammatory response such as TNF-a, IL-la, IL-Ιβ, and IL- 2- IL18. The at least one capture antibody, or functional portion thereof, can effectively bind and remove one or more of TNF-a, IL-la, IL-Ιβ, IL-2- IL18, or other cytokines involved in inflammation.

[0026] In some embodiments utilizing the sequentially connected housings, each housing will contain a unique filtration matrix distinct from the filtration matrices of the other sequentially connected housings. Such configuration allows for in situ customizability for individual patients and individual inflammation responses to only target specified cytokines for removal.

[0027] In some embodiments utilizing the sequentially connected housings, some housings may contain a unique filtration matrix distinct from the filtration matrices of the other sequentially connected housings, while others contain the same filtration matrix. Such configuration allows for in situ customizability for individual patients and individual inflammation responses to only target a therapeutically effective amount of specified cytokines for removal.

[0028] In alternative embodiments, a peripheral blood filtration device is provided that includes a housing configured to receive peripheral blood through a blood introducing port, contacting peripheral blood with a filtration matrix, and a peripheral blood outflow channel, wherein: the filtration matrix is in situ configured and assembled for individual patients and individual inflammation responses to only target a therapeutically effective amount of specified cytokines for removal.

BRIEF DESCRIPTION OF DRAWINGS

[0029] FIG. 1 is an illustration of a configuration of the extracorporeal peripheral blood filtration device according to an exemplary embodiment of the invention, wherein the filtration matrix comprises a capture antibody immobilized on a plurality of beads.

[0030] FIG. 2 is an illustration of a representative method utilizing sequentially connected housings of the extracorporeal peripheral blood filtration device according to an exemplary embodiment of the invention.

[0031] FIG. 3 is an illustration of a configuration of the extracorporeal peripheral blood filtration device according to an exemplary embodiment of the invention, wherein the filtration matrix comprises a capture antibody immobilized on a strip.

[0032] FIG. 4 is an illustration of a configuration of a single bead, a plurality of which comprises the filtration matrix of the extracorporeal peripheral blood filtration device according to an exemplary embodiment of the invention.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0033] The present invention may be understood more readily by reference to the following detailed description of the preferred embodiments of the invention. However, although different components and methods are disclosed and described, it is to be understood that this invention is not limited to specific formulations, assemblies or configurations, conditions, or methods, as such may vary, and any modifications thereto and variations therein will be apparent to those skilled in the art. It is also to be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to be limiting.

I. Definitions

[0034] It must be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the," include plural forms unless the context clearly indicates otherwise. Thus, for example, reference to "a material" includes one or more of such same or different materials, and reference to "the method" includes reference to equivalent steps and methods known to those of ordinary skill in the art that could be modified or substituted for the methods described herein.

[0035] As used herein, the term "antigen" refers to a target molecule that is specifically bound by an antibody through its antigen recognition site. The antigen may be monovalent or polyvalent, i.e., it may have one or more epitopes recognized by one or more antibodies. Examples of kinds of antigens that can be recognized by antibodies include polypeptides, oligosaccharides, glycoproteins, polynucleotides, lipids, etc.

[0036] As used herein, the term "capture antibody" means an antibody, or a functional portion or fragment thereof, with a high binding affinity for cytokines involved in an inflammation response, preferably a protein with a high binding affinity for only one cytokine, including (but not limited to) TNF-a, IL-la, IL-Ιβ, and IL-2- IL18. The term "capture antibody" is used in the broadest sense and includes polyclonal and monoclonal antibodies, including intact antibodies and functional (antigen-binding) antibody fragments, including fragment antigen binding (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rlgG) fragments, single chain antibody fragments, including single chain variable fragments (scFv), and single domain antibodies (e.g., sdAb, sdFv, nanobody) fragments. The term encompasses genetically engineered and/or otherwise modified forms of immunoglobulins, such as intrabodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecific, e.g., bispecific, antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFv, tandem tri-scFv. Unless otherwise stated, the term "capture antibody" should be understood to encompass functional antibody fragments thereof. The term also encompasses intact or full-length antibodies, including antibodies of any class or subclass, including IgG and sub-classes thereof, IgM, IgE, IgA, and IgD.

[0037] The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0038] Among the provided antibodies are antibody fragments. An "antibody fragment" refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds the antigen to which the intact antibody binds. Examples of antibody fragments include but are not limited to Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g. scFv); and multispecific antibodies formed from antibody fragments. In particular embodiments, the antibodies are single-chain antibody fragments comprising a variable heavy chain region and/or a variable light chain region, such as scFvs.

[0039] Single-domain antibodies are antibody fragments comprising all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody. In certain embodiments, a single-domain antibody is a camelid single-domain antibody.

[0040] Antibody fragments can be made by various techniques, including but not limited to proteolytic digestion of an intact antibody as well as production by recombinant host cells. In some embodiments, the antibodies are recombinantly-produced fragments, such as fragments comprising arrangements that do not occur naturally, such as those with two or more antibody regions or chains joined by synthetic linkers, e.g., peptide linkers, and/or that are may not be produced by enzyme digestion of a naturally-occurring intact antibody.

[0041] A "humanized" antibody is an antibody in which all or substantially all

CDR amino acid residues are derived from non-human CDRs and all or substantially all FR amino acid residues are derived from human FRs. The term "chimeric" antibody refers to an antibody in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.

[0042] Among the provided antibodies are monoclonal antibodies, including monoclonal antibody fragments. The term "monoclonal antibody" as used herein refers to an antibody obtained from or within a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical, except for possible variants containing naturally occurring mutations or arising during production of a monoclonal antibody preparation, such variants generally being present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different epitopes, each monoclonal antibody of a monoclonal antibody preparation is directed against a single epitope on an antigen. The term is not to be construed as requiring production of the antibody by any particular method. A monoclonal antibody may be made by a variety of techniques, including but not limited to generation from a hybridoma, recombinant DNA methods, phage-display and other antibody display methods.

[0043] As used herein, the term "specific binding" refers to the specificity of a binding reagent, e.g., an antibody, such that it preferentially binds to a target antigen. Recognition by a binding reagent or an antibody of a particular target in the presence of other potential interfering substances is one characteristic of such binding. Preferably, binding reagents, antibodies or antibody fragments that are specific for or bind specifically to a target antigen bind to the target antigen with higher affinity than binding to other non- target substances. Also preferably, binding reagents, antibodies or antibody fragments that are specific for or bind specifically to a target antigen avoid binding to a significant percentage of non-target substances, e.g., non-target substances present in a testing sample. In some embodiments, binding reagents, antibodies or antibody fragments of the present disclosure avoid binding greater than about 90% of non-target substances, although higher percentages are clearly contemplated and preferred. For example, binding reagents, antibodies or antibody fragments of the present disclosure avoid binding about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, and about 99% or more of non-target substances. In other embodiments, binding reagents, antibodies or antibody fragments of the present disclosure avoid binding greater than about 10%, 20%, 30%, 40%, 50%, 60%, or 70%, or greater than about 75%, or greater than about 80%, or greater than about 85% of non-target substances.

[0044] "Beads" or "capture beads" as used herein refer to particles whereupon desired capture antibodies have been immobilized. The beads generally are uniform in size within a single filtration matrix, but may vary in size ranging from about 1 nm to about 10,000 nm between different filtration matrices. The preferred shape is spherical; however, particles of any other shape can be employed since this parameter is immaterial to the nature of the invention.

[0045] "Strips" or "capture strips" as used herein refer to elongated flat elements whereupon desired capture beads or desired capture antibodies have been immobilized. The strips generally are thin films uniform in size, but may vary in size and color depending on the amount and type of capture antibody immobilized. A preferred shape is a rectangular prism; however, elements of any other shape can be employed since this parameter is immaterial to the nature of the invention.

[0046] "High surface area configuration or matrix" as used herein refers to non- porous elements with flat sides whereupon desired capture beads or desired capture antibodies have been immobilized. Generally high surface area configuration or matrix refers to elements having a high surface area to volume ratio. A preferred shape comprises hollow elements such as tubes; however, elements of any other shape can be employed since this parameter is immaterial to the nature of the invention.

[0047] "Filtration matrix" as used herein in some embodiments refer to a composition comprising a plurality of one or more types of beads whereupon one or more capture antibodies have been immobilized. In other embodiments, filtration matrix refers to a composition comprising one or more strips whereupon one or more capture antibodies have been immobilized. If the filtration matrix comprises a plurality of strips, the strips may be of either the same or of different types. In yet other embodiments, filtration matrix refers to any high surface area configuration or matrix whereupon one or more capture antibodies have been immobilized.

[0048] As used herein, "an effective amount" refers to the amount sufficient to bring about a desired result in an experimental setting. A "therapeutically effective amount" refers to an amount sufficient to produce a therapeutic response or beneficial clinical result in a patient.

[0049] As used herein, the terms "patient" and "individual" refer to any person or other subject that is at risk of or suffering from a disordered inflammation response. It is envisioned that the "patient" may also be a non-human animal, such as in veterinary applications of the invention.

II. Extracorporeal Blood Filtration Device

[0050] The device can comprise a stand-alone, or unitary, extracorporeal component that can be coupled in-line to blood tubing at time of use. In this arrangement (see FIG. 1), the device in its most basic form includes a housing 1, wherein: the housing is a flow-through vessel, such as a column or funnel wherein the peripheral blood is introduced at one end or orifice, and a sample is eluted from another end or orifice. The housing 1 contains a filtration matrix 2 that removes cytokines by high-affinity binding. In an illustrative embodiment the filtration matrix includes a plurality of beads 3 whereupon at least one capture antibody 4 is immobilized (FIG. 1). In other embodiments, the filtration matrix includes one or more strips 8 whereupon a plurality of beads 3 with at least one capture antibody 4 are immobilized (FIG. 3). In yet other embodiments, the filtration matrix includes one or more strips 8 whereupon at least one capture antibody 4 is directly immobilized.

[0051] The housing 1 includes a blood introducing port 5 that can be connected to blood tubing 7, for directing peripheral blood into the housing 1 to come in contact with the filtration matrix 2. The housing 1 also includes an outflow channel 6 that can be connected to blood tubing 7, for directing the peripheral blood from the housing 1 after contact with the filtration matrix 2, during which all or a portion of the targeted cytokines present are removed.

[0052] The transport of the peripheral blood through the filtration matrix 2 can be accomplished in various ways. When used as a stand-alone device, in one embodiment, an external pump can be used to convey the peripheral blood from the patient, through the device, and back to the patient. In this arrangement, blood tubing 7 connected to the blood introducing port 5 can be coupled via a suitable blood access to an artery, while blood tubing 7 connected to the outflow channel 6 can be coupled by a suitable blood access to a vein.

[0053] When used in association with other peripheral blood filtration or dialysis systems, the introducing port 5 and the outflow channel 6 can be configured to comprise an exchangeable component than can be directly coupled to conventional intravenous blood access catheters or be configured with fittings matching the system, such that the device can be connected in-line with the blood tubing already used by the system. In one embodiment the introduction port 5 and the out flow channel 6 of the device shown in FIG. 1 can include conventional mating luer fittings to enable quick attachment and removal of the device from the in-line blood flow route.

[0054] In other embodiments, peripheral blood is taken from the patient and filtered using the device as stand-alone, or unitary, extracorporeal component in a separate and distinct system not connected to the patient. In this arrangement, the peripheral blood is conveyed through the device, and the filtered blood is collected for immediate or later reinfusion back into the patient.

[0055] In an alternative embodiment shown in FIG. 2, two or more housings 1 can be sequentially connected in-line to enable the customizability of both which cytokines and the amount of cytokines being targeted for removal.

[0056] In another alternative embodiment, customizability of both which cytokines and the amount of cytokines being targeted for removal can be achieved by configuring and assembling the housing 1 to contain a filtration matrix 2 that only targets a therapeutically effective amount of specified cytokines for removal.

III. Filtration Matrix

[0057] The filtration matrix can be variously constructed. In a preferred embodiment, the filtration matrix comprises a column of beads coated with antibodies specific for binding cytokines. A column design to ensure homogenous distribution of peripheral blood over the filtration matrix is also preferred to prevent clogging and formation of channels and short circuits. [0058] In an illustrative embodiment (FIG. 1), the filtration matrix consists of a plurality of spherical beads whereupon at least one type of capture antibody with a high binding affinity for at least one cytokine has been immobilized.

[0059] In other embodiments, the filtration matrix comprises a plurality of beads whereupon two or more specific capture antibodies are immobilized.

[0060] In other embodiments, the filtration matrix comprises a mixture of two or more pluralities of beads with each plurality of beads having a unique immobilized capture antibody.

[0061] In another illustrative embodiment (FIG. 3), the filtration matrix consists of a strip whereupon a plurality of beads with at least one type of capture antibody with a high binding affinity for at least one cytokine have been immobilized.

[0062] In other embodiments, the filtration matrix comprises a strip whereupon a plurality of beads with two or more specific capture antibodies are immobilized.

[0063] In other embodiments, the filtration matrix comprises a strip with an immobilized mixture of two or more pluralities of beads with each plurality of beads having a unique immobilized capture antibody.

[0064] In other embodiments, the filtration matrix consists of a strip or other high surface area configuration or matrix whereupon one or more capture antibodies with a high binding affinity for at least one cytokine have been directly immobilized.

[0065] In still other embodiments, the filtration matrix comprises a strip or other high surface area configuration or matrix whereupon two or more specific capture antibodies are immobilized.

IV. Capture Beads

[0066] An illustrative embodiment of the structure/configuration of a capture bead 3 of the invention is depicted in FIG. 4. Generally, each bead is coated with one or more specific antibodies.

[0067] Beads can be either solid or hollow and be made of any biocompatible material such as, but not limited to, metals like: titanium, stainless steels, cobalt and magnesium alloys, ceramics like: hydroxyapatite and phosphate tricalcium, and polymers like: polyacrylonitrile, polysulfone, polyethersulfone, polyethylene, polymethylmethacrylate, polytetrafluoroethylene, polyester, polypropylene, polyether ether ketone, Nylon, polyether-block co-polyamide polymers, polyurethanes such as aliphatic polyether polyurethanes, polyvinyl chloride, thermoplastic, fluorinated ethylene propylene, cellulose, collagen, silicone or any combination thereof.

[0068] Capture antibodies may be polyclonal or monoclonal cytokine antibodies or receptors, fragments derived therefrom, or recombinant constructs having the binding activity of such antibodies or receptors.

[0069] The antibodies, or functional interleukin binding portions thereof, employed in the present invention can be immobilized on any appropriate material by any appropriate technique. Many methods of immobilizing antibodies are known in the art. Such methods include, but not limited to covalent coupling, direct adsorption, physical entrapment, and attachment to a protein-coated surface. Any combination of bead and binding technique which leaves the capture antibody immunoreactive, yet sufficiently immobilizes the antibody so that it can be retained with any bound cytokine during filtration, can be employed in the present invention.

[0070] In one representative embodiment, a recombinant capture antibody mimicking the ligand binding region of the high affinity cell- surface receptors on the target cells for a specific cytokine is immobilized on the surface of a bead by first coating it with a water-insoluble protein such as zein, collagen, fibrinogen, keratin, or glutelin. The capture antibody is then attached by simply contacting the protein-coated surface with an aqueous solution of the antibody and allowing it to dry.

[0071] In alternative embodiments, the capture beads are bound to a strip (FIG. 3).

V. Capture Strips [0072] In embodiments capture strips are coated with one or more specific antibodies. In some embodiments, capture strips are elongated elements.

[0073] The strips can be either solid or hollow and be made of any biocompatible material such as, but not limited to, metals like: titanium, stainless steels, cobalt and magnesium alloys, ceramics like: hydroxyapatite and phosphate tricalcium, and polymers like: polyacrylonitrile, polysulfone, polyethersulfone, polyethylene, polymethylmethacrylate, polytetrafluoroethylene, polyester, polypropylene, polyether ether ketone, Nylon, polyether-block co-polyamide polymers, polyurethanes such as aliphatic polyether polyurethanes, polyvinyl chloride, thermoplastic, fluorinated ethylene propylene, cellulose, collagen, silicone or any combination thereof.

[0074] Capture antibodies may be polyclonal or monoclonal cytokine antibodies or receptors, fragments derived therefrom, or recombinant constructs having the binding activity of such antibodies or receptors.

[0075] The antibodies, or functional interleukin binding portions thereof, employed in the present invention can be immobilized on any appropriate material by any appropriate technique. Many methods of immobilizing antibodies are known in the art. Such methods include, but not limited to covalent coupling, direct adsorption, physical entrapment, and attachment to a protein-coated surface. Any combination of strip and binding technique which leaves the capture antibody immunoreactive, yet sufficiently immobilizes the antibody so that it can be retained with any bound cytokine during filtration, can be employed in the present invention.

[0076] In one representative embodiment, a recombinant capture antibody mimicking the ligand binding region of the high affinity cell- surface receptors on the target cells for a specific cytokine is immobilized on the surface of a strip by first coating it with a water-insoluble protein such as zein, collagen, fibrinogen, keratin, or glutelin. The capture antibody is then attached by simply contacting the protein-coated surface with an aqueous solution of the antibody and allowing it to dry.

[0077] These and other embodiments of the invention will be apparent to the skilled artisan in view of the present disclosure.

Claims

CLAIMS WHAT IS CLAIMED IS:

1. An extracorporeal peripheral blood filtration device, comprising: a housing comprising a blood introducing port and a blood outflow channel; wherein the housing contains a filtration matrix that when contacted with blood specifically removes inflammation-regulating cytokines.

2. The device of claim 1 , wherein the device is configured to receive blood through a flow route compatible with dialysis or other peripheral blood filtration systems.

3. The device of claim 1, wherein the device is configured to receive blood through a flow route as part of dialysis or other peripheral blood filtration methods.

4. The device of claim 1 , wherein the filtration matrix comprises at least one capture antibody immobilized on a plurality of beads, on one or more strips or other high surface area embedding matrix.

5. The device of claim 1, wherein the filtration matrix comprises at least two capture beads, wherein each capture bead comprises at least one antibody specific for a cytokine immobilized on a plurality of beads, and wherein said at least two capture beads are specific for different types of inflammation regulating cytokines.

6. The device of claim 1 , wherein the filtration matrix comprises at least two capture strips, wherein each capture strip comprises at least one antibody specific for a cytokine immobilized on one or more strips, and wherein said at least two capture strips are specific for different types of inflammation regulating cytokines.

7. The device of claim 1, wherein the filtration matrix comprises at least two capture elements, wherein each capture element comprises at least one antibody specific for a cytokine immobilized on a high surface area configuration or matrix, and wherein said at least two capture elements are specific for different types of inflammation regulating cytokines.

8. The device of claim 4, wherein the at least one capture antibody has a high affinity for at least one inflammation regulating cytokine.

9. The device of claim 4, wherein the at least one capture antibody has a high affinity for one or more cytokines but not limited to selection from a group consisting of TNF-a, IL-la, IL-Ιβ, and IL-2- IL18.

10. The device of claim 1, wherein the filtration matrix is configured to target a therapeutically effective amount of cytokines, identified as being overexpressed, for removal.

11. A method for treating an inflammatory disease or condition in a patient comprising: providing the extracorporeal peripheral blood filtration device of claim 1 , and removing at least one inflammation regulating cytokine from the peripheral blood by contacting said peripheral blood with the filtration matrix by passing said peripheral blood through the device.

12. The method of claim 11, wherein the extracorporeal peripheral blood filtration device is configured to receive blood through a flow route compatible with dialysis or other peripheral blood filtration systems.

13. The method of claim 11, wherein the filtration matrix comprises at least one capture antibody immobilized on a plurality of beads, on one or more strips or other high surface area embedding matrix.

14. The method of claim 11, wherein the filtration matrix comprises at least two capture beads, wherein each capture bead comprises at least one antibody specific for a cytokine immobilized on a plurality of beads, and wherein said at least two capture beads are specific for different types of inflammation causing cytokines.

15. The method of claim 11, wherein the filtration matrix comprises at least two capture strips, wherein each capture strip comprises at least one antibody specific for a cytokine immobilized on one or more strips, and wherein said at least two capture strips are specific for different types of inflammation causing cytokines.

16. The method of claim 11, wherein the filtration matrix comprises at least two capture elements, wherein each capture element comprises at least one antibody specific for a cytokine immobilized on a high surface area configuration or matrix, and wherein said at least two capture elements are specific for different types of inflammation causing cytokines.

17. The method of claim 11, wherein the at least one capture antibody has a high affinity for one or more cytokines but not limited to selection from a group consisting of TNF-a, IL-la, IL-Ιβ, and IL-2- IL18.

18. The method of claim 11, wherein peripheral blood from the patient is conveyed through the provided extracorporeal peripheral blood filtration device, at least one inflammation regulating cytokine is removed from the peripheral blood by contacting said peripheral blood with the filtration matrix by passing said peripheral blood through the device, and the peripheral blood is directly reinfused into said patient.

19. The method of claim 11, wherein the method further comprises the following steps; (a) taking peripheral blood from the patient; (b) conveying said peripheral blood through the provided extracorporeal peripheral blood filtration device, wherein the extracorporeal peripheral blood filtration device is part of an isolated blood filtration system; (c) monitoring the presence and amount of cytokines in the filtered blood as it exits from said extracorporeal peripheral blood filtration device, and (d) repeating steps (b) and (c) until desired cytokine concentrations have been achieved whereupon the filtered blood is reinfused into said patient. A method for treating an inflammatory disease or condition in a patient comprising: identifying a patient experiencing a disordered inflammatory response, identifying cytokines being overexpressed, determining concentrations of overexpressed cytokines, configuring and assembling the extracorporeal peripheral blood filtration device of claim 1, wherein the filtration matrix is configured to target a therapeutically effective amount of identified overexpressed cytokines for removal, providing the assembled extracorporeal peripheral blood filtration device, and removing the targeted overexpressed inflammation regulating cytokines from the peripheral blood by contacting said peripheral blood with the filtration matrix by passing said peripheral blood through the device.