System and method for removal of immune inhibitors from biological fluids

Abstract

The present system and method are useful for the removal of immune inhibitors such as soluble TNF receptors from the body fluid of cancer patients. In some embodiments, soluble TNF-Receptors 1 and 2 are selectively removed from plasma at 80% or more efficiency. In some embodiments, the system includes an immobilized capture ligand of a single chain TNFα. The system and method are useful for the treatment of different cancer types, stages and severity.

What is claimed is: 
     
         1 . A system for removing at least one target component of body fluid, the system comprising:
 an inlet configured to receive the body fluid from a patient;   a sequestering chamber coupled to the inlet and configured to receive the body fluid from the inlet, the sequestering chamber comprising:
 a capture support configured to bind to the at least one target component of the body fluid to capture the at least one target component in the sequestering chamber responsive to contact between the capture support and the body fluid, the capture support configured to bind to the at least one target component to reduce an amount of the at least one target component in the body fluid; and 
 first and second access ports configured to provide access to the sequestering chamber separate from the inlet, the first and second access ports configured to facilitate insertion and/or removal of the capture support to and/or from the sequestering chamber; 
   an outlet configured to pass the body fluid having the reduced amount of the at least one target component from the sequestering chamber for optional reintroduction of some or all of the body fluid having the reduced amount of the at least one target component back into the patient; and   one or more filters configured to separate the capture support in the sequestering chamber from the inlet and the outlet, the one or more filters configured to retain the capture support within the sequestering chamber.   
     
     
         2 . The system of  claim 1 , wherein (a) a capture efficiency of the capture support binding to the at least one target component is 80% or more at a flow rate of 45 mL per minute of plasma flow or less, and optionally wherein
 (b) a binding affinity of the capture support to the at least one target component is at least about 10 −7 K D  and/or   (c) a leach rate of the capture support through the outlet is less than about 100 ng/m L/min.   
     
     
         3 . The system of  claim 1 , wherein (b) a binding affinity of the capture support to the at least one target component is 10 −7  K D  or greater, and optionally wherein
 (a) a capture efficiency of the capture support binding to the at least one target component is 80% or more at a flow rate of 45 mL/min or less, and/or   (c) a leach rate of the capture support through the outlet is less than about 100 ng/m L/min.   
     
     
         4 . The system of  claim 1 , wherein (c) a leach rate of the capture support through the outlet is less than about 100 ng/mL/min, and optionally wherein
 (a) a capture efficiency of the capture support binding to the at least one target component is 80% or more at a flow rate of 45 mL/min or less, and/or   (b) a binding affinity of the capture support to the at least one target component is 10 −7  K D  or greater.   
     
     
         5 . The system of any of  claims 1 - 4 , wherein the body fluid comprises plasma. 
     
     
         6 . The system of any of  claims 1 - 5 , wherein the at least one target component comprises a protein, complex, assembly, or cell. 
     
     
         7 . The system of any of  claims 1 - 6 , wherein the at least one target component comprises one or more plasma components that function to inhibit anti-cancer immune responses in the patient. 
     
     
         8 . The system of any of  claims 1 - 7 , wherein the at least one target component comprises one or more immune inhibitors. 
     
     
         9 . The system of any of  claims 1 - 8 , wherein the at least one target component comprises a soluble TNFα receptor. 
     
     
         10 . The system of any of  claims 1 - 9 , wherein the at least one target component comprises an sTNF-R1 receptor and/or an sTNF-R2 receptor. 
     
     
         11 . The system of  claims 1 - 10 , wherein the capture support comprises an affinity chromatography support material, hollow fiber membranes, sheet membranes, membrane cassettes, beads, or rolled sheet membranes. 
     
     
         12 . The system of  claim 11 , wherein the capture support comprises the affinity chromatography support material, and wherein the affinity chromatography support material comprises sepharose, agarose, or acrylamide. 
     
     
         13 . The system of any of  claims 1 - 12 , wherein the capture support comprises a porous or non-porous matrix material. 
     
     
         14 . The system of any of  claims 1 - 13 , wherein the capture support is configured to bind to more than one target component of the body fluid. 
     
     
         15 . The system of any of  claims 1 - 14 , wherein the capture support comprises a solid support having antibodies, antibody fragments, binding peptides, aptamers, or avimers immobilized thereon. 
     
     
         16 . The system of  claim 15 , wherein the antibodies are selected from the group consisting of IgA, IgD, IgE, IgG, IgM, and combinations thereof. 
     
     
         17 . The system of any of  claims 1 - 16 , wherein the capture support comprises TNFα, multimers of TNFα, single chain TNFα, fragments of TNFα, multimers of fragments of TNFα, or combinations thereof. 
     
     
         18 . The system of any of  claims 1 - 17 , wherein the capture support comprises an sc-TNFα ligand, optionally the sequence or partial sequence of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. 
     
     
         19 . The system of  claim 18 , wherein the capture support comprises a trimeric form of the sc-TNFα ligand. 
     
     
         20 . The system of  claim 19 , wherein the trimeric form of the sc-TNFα ligand comprises the sequence or partial sequence of SEQ ID NO:2 or SEQ ID NO:3. 
     
     
         21 . The system of any of  claims 1 - 20 , wherein the capture support comprises ligands bound to beads. 
     
     
         22 . The system of  claim 21 , wherein the ligands have a given density and orientation on a given bead, the density and orientation configured to enhance binding between the ligands and the at least one target component of the body fluid. 
     
     
         23 . The system of any of  claim 21  or  22 , wherein a size, number, density, and/or concentration of the beads is configured to facilitate a laminar flow of the body fluid through the beads to enhance the binding between the ligands and the at least one target component of the body fluid. 
     
     
         24 . The system of any of  claims 21 - 23 , wherein the beads are quenched in ethanolamine to enhance binding specificity. 
     
     
         25 . The system of any of  claims 1 - 24 , wherein the body fluid is whole blood. 
     
     
         26 . The system of any of  claims 1 - 25 , wherein the inlet, the sequestering chamber, and the outlet form an extracorporeal closed-circuit column. 
     
     
         27 . The system of  claim 26 , wherein the extracorporeal closed-circuit column is configured to remain sterile during operation. 
     
     
         28 . The system of any of  claim 26  or  27 , further comprising a target component outlet port configured to facilitate sampling or removal of all or part of the captured at least one target component without compromising the extracorporeal closed-circuit column. 
     
     
         29 . The system of any of  claims 26 - 28 , further comprising an elution reagent port configured to facilitate introduction of an elution reagent into the sequestering chamber without compromising the extracorporeal closed-circuit column. 
     
     
         30 . The system of  claim 29 , wherein the elution reagent port is further configured to receive a conditioning agent configured to prepare the system for reuse. 
     
     
         31 . The system of any of  claims 1 - 30 , further comprising a pump configured to drive a reconditioning agent through the inlet, the sequestering chamber, and the outlet. 
     
     
         32 . The system of  claim 31 , wherein the pump comprises a syringe pump, a peristaltic pump, a piston pump, a diaphragm pump, or a combination thereof. 
     
     
         33 . The system of any of  claims 1 - 32 , wherein the one or more filters have an average pore diameter between 3 microns and 100 microns. 
     
     
         34 . The system of any of  claims 1 - 33 , further comprising one or more additional sequestering chambers including capture supports having the same functionality. 
     
     
         35 . The system of  claim 34 , wherein the one or more additional sequestering chambers combine with the sequestering chamber to form a multistage separation circuit configured to bind with a plurality of different target components. 
     
     
         36 . The system of any of  claims 1 - 35 , wherein the patient is a human or a veterinary subject. 
     
     
         37 . A method for removing the at least one target component of the body fluid with the system of any of  claims 1 - 36 , the method comprising:
 conducting the body fluid from the patient through the inlet to the sequestering chamber;   binding the at least one target component of the body fluid to capture the at least one target component in the sequestering chamber to reduce the amount of the at least one target component in the body fluid; and optionally   passing some or all of the body fluid having the reduced amount of the at least one target component from the sequestering chamber through the outlet for reintroduction back into the patient.   
     
     
         38 . The method of  claim 37 , further comprising measuring the reduced amount of the at least one target component in the body fluid reintroduced back into the patient. 
     
     
         39 . The method of  claim 38 , wherein the measuring comprises one or more of LC-MS, HPLC, UHPLC, resistance measurements, light emission measurements, chemiluminescence, electroluminescence, electrochemiluminescence, chromatographic monitoring, positron emission tomography (PET), x-ray computed tomography (CT), magnetic resonance imaging (MRI), ultrasound, gamma camera, single photon emission computed tomography (SPECT), ELISA, SPR, or BLI. 
     
     
         40 . The method of any of  claims 37 - 39 , further comprising measuring a leach rate of the capture support in the body fluid reintroduced back into the patient. 
     
     
         41 . The method of any of  claims 37 - 40 , wherein the method is for human or veterinary use.