Devices and methods of cell capture and analysis

Abstract

The present invention provides a device for isolating target biomolecules or cells from samples, particularly biological samples. In particular, the device comprises a loading mixture, which contains the biological sample and a first binding entity that specifically binds to the target biomolecule or target cell; and a micro-channel coated with a second binding entity that binds directly or indirectly to the first binding entity. Methods of capturing, detecting, and/or evaluating target biomolecules or target cells (e.g. cancer cells) in biological samples are also disclosed.

1 . A method for characterizing a tumor cell from a biological sample that expresses target entity, the method comprising:
 a) combining said biological sample with a first binding entity that selectively binds to said target entity to form bound tumor cells, wherein said first binding entity further comprises a third binding entity and a capturable moiety,   b) contacting said first binding entity bound tumor cells with a surface, wherein said surface comprises a second binding entity, wherein said third binding entity capturable moiety binds to said second binding entity on said surface to form a tumor cell bound surface,   c) contacting said tumor cell bound surface with a detectible entity that binds said third binding moiety and   d) identifying and characterizing said tumor cell on said tumor cell bound surface, wherein said characterizing comprises enumerating said tumor cell, determining an expression level for said target entity or any combination thereof.   
     
     
         2 . The method according to  claim 1 , wherein said capture moiety is selected from the group consisting of biotin, avidin, streptavidin, digoxigenin, anti-digoxigenin Ab, or a derivative thereof. 
     
     
         3 . The method according to  claim 1 , wherein said second binding entity comprises biotin or a derivative thereof and said capture moiety comprises streptavidin or a derivative thereof. 
     
     
         4 . The method according to the method of  claim 1 , wherein said third binding moiety and said capturable moiety are the same moiety. 
     
     
         5 . The method according to  claim 1 , wherein said third binding moiety and said capturable moiety are biotin. 
     
     
         6 . The method according to  claim 1 , wherein said biological sample is a physiological fluid or tissue selected from the group consisting of blood, plasma, bone marrow, serum, semen, vaginal secretions, urine, saliva, amniotic fluid, cerebral spinal fluid, synovial fluid, lung lavages, fine needle aspirates (FNA) and biopsy tissue sample. 
     
     
         7 . The method according to  claim 1 , wherein said tumor cell is a circulating tumor cell. 
     
     
         8 . The method according to  claim 1 , wherein said tumor cell is a cancer cell selected from a breast cancer cell, a prostate cancer cell, a colorectal cancer cell, a lung cancer cell, a pancreatic cancer cell, a ovarian cancer cell, a bladder cancer cell, a endometrial cancer cell, a cervical cancer cell, a liver cancer cell, a renal cancer cell, a thyroid cancer cell, a bone cancer cell, a lymphoma cancer cell, a melanoma cancer cell, and a non-melanoma cancer cell. 
     
     
         9 . The method according to  claim 1 , wherein the tumor cell is:
 a) a breast cancer cell and wherein said target that binds the actionable antigen is an antibody that specifically binds to Her2/neu, EpCAM, MUC-1, EGFR, TAG-12, IGF1R, TACSTD2, CD318, CD104, or N-cadherin, or said target is an antibody cocktail specifically binding a combination thereof;   b) a melanoma cancer cell and wherein said target that binds the actionable antigen is an antibody that specifically binds to melanocyte differentiation antigens, oncofetal antigens, tumor specific antigens, SEREX antigens or a combination thereof;   c) a prostate cancer cell and wherein said target that binds the actionable antigen is an antibody that specifically binds to EpCAM, MUC-1, EGFR, PSMA, PSA, TACSTD2, PSCA, PCSA, CD318, CD104, or N-cadherin, or said target is an antibody cocktail specifically binding a combination thereof;   d) a colorectal cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to EpCAM, CD66c, CD66e, CEA, TACSTD2, CK20, CD104, MUC-1, CD318, or N-cadherin, or said target is an antibody cocktail specifically binding a combination thereof;   e) a lung cancer cell and said target that binds said actionable antigen is an antibody that specifically binds to CK18, CK19, TACSTD2, CD318, CD104, CEA, EGFR, or EpCAM, or said target is an antibody cocktail specifically binding a combination thereof;   f) a pancreatic cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to MUC-1, CEA, TACSTD2, CD104, CD318, N-cadherin, MUC-1, or EpCAM, or said target is an antibody cocktail specifically binding a combination thereof;   g) an ovarian cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to MUC-1, TACSTD2, CEA, CD318, CD104, N-cadherin, or EpCAM, or said target is an antibody cocktail specifically binding a combination thereof;   h) an endothelial bladder cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to CD34, CD146, CD62, CD105, CD106, VEGF receptor, or MUC-1, or said target is an antibody cocktail specifically binding a combination thereof;   i) an epithelial bladder cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to TACSTD2, EpCAM, CD318, EGFR, 6B5, N-cadherin or folate binding receptor, or said target is an antibody cocktail that specifically binds a combination thereof;   j) a cancer stem cell and said target that binds the actionable antigen is an antibody that specifically binds to CD133, CD135, CD117, or CD34, or said target is an antibody cocktail that specifically binds a combination thereof;   k) a circulating cancer cell that expresses mesenchymal antigens and said target that binds the actionable antigen is an antibody that specifically binds to FGFR1, FGFR4, EGFR, folate binding receptor, N-cadherin or MSC, or said target is an antibody that specifically binds a combination thereof; and/or   l) a circulating cancer cell that expresses angiogenesis surface antigens and said target that binds the actionable antigen is an antibody that specifically binds to a VEGF receptor.   
     
     
         10 . The method according to  claim 1 , wherein said surface is a microchannel. 
     
     
         11 . The method according to  claim 1 , wherein said cancer or inflammatory disease treatment is cancer or inflammatory disease therapeutics development, a cancer or inflammatory disease therapy decision, cancer or inflammatory disease therapy monitoring, determination of cancer or inflammatory disease progression, or a minimal residual cancer or inflammatory disease determination. 
     
     
         12 . A method for characterizing tumor cells in a biological fluid expressing a target entity for cancer or inflammatory disease treatment, said method comprising:
 a) combining said biological fluid with an antibody that selectively binds said target entity, wherein said antibody comprises biotin to form antibody bound tumor cells,   b) contacting said antibody bound tumor cells with a surface, wherein said surface comprises streptavidin, to form a tumor cell bound surface,   c) contacting said tumor cell bound surface with a streptavidin fluorophore to form labeled tumor cells, and   d) characterizing said labeled tumors cells, wherein said characterizing comprises enumerating said labeled tumor cells, determining an expression level for said target entity, determining a signal localization for said actionable antigen with respect to said tumor cells, determining size and/or morphology of said tumor cells, determining a drug treatment biomarker associated with said tumor cells, or any combination thereof.   
     
     
         13 . A method for characterizing a tumor cell from a biological sample that expresses target entity, the method comprising:
 a) combining said biological sample with a first antibody that selectively binds to said target entity to form antibody bound tumor cells,   b) incubating said antibody bound tumor cells with a biotinylated secondary antibody that targets the first antibody to form antibody complexed tumor cells,   c) contacting said antibody complexed tumor cells with a surface, wherein said surface contains avidin or streptavidin to form a tumor cell bound surface,   d) contacting said tumor cell bound surface with a streptavidin or avidin bound reporter group that binds said biotinylated secondary antibody, and   e) identifying and or characterizing said tumor cell on said tumor cell bound surface, wherein said characterizing comprises enumerating said tumor cell, determining an expression level for said target entity, detecting a drug treatment biomarker associated with said tumor cell, or any combination thereof.   
     
     
         14 . The method according to  claim 13 , wherein said tumor cell is a tumor cell in a physiological fluid. 
     
     
         15 . The method according to  claim 13 , wherein said biological sample is a physiological fluid or tissue selected from the group consisting of blood, plasma, bone marrow, serum, semen, vaginal secretions, urine, saliva, amniotic fluid, cerebral spinal fluid, synovial fluid, lung lavages, fine needle aspirates (FNA) and biopsy tissue sample. 
     
     
         16 . The method according to  claim 13 , wherein said tumor cell is a cancer cell selected from a breast cancer cell, a prostate cancer cell, a colorectal cancer cell, a lung cancer cell, a pancreatic cancer cell, a ovarian cancer cell, a bladder cancer cell, a endometrial cancer cell, a cervical cancer cell, a liver cancer cell, a renal cancer cell, a thyroid cancer cell, a bone cancer cell, a lymphoma cancer cell, a melanoma cancer cell, and a non-melanoma cancer cell. 
     
     
         17 . The method according to  claim 13 , wherein the tumor cell is:
 a) a breast cancer cell and wherein said target that binds the actionable antigen is an antibody that specifically binds to Her2/neu, EpCAM, MUC-1, EGFR, TAG-12, IGF1R, TACSTD2, CD318, CD104, or N-cadherin, or said target is an antibody cocktail specifically binding a combination thereof;   b) a melanoma cancer cell and wherein said target that binds the actionable antigen is an antibody that specifically binds to melanocyte differentiation antigens, oncofetal antigens, tumor specific antigens, SEREX antigens or a combination thereof;   c) a prostate cancer cell and wherein said target that binds the actionable antigen is an antibody that specifically binds to EpCAM, MUC-1, EGFR, PSMA, PSA, TACSTD2, PSCA, PCSA, CD318, CD104, or N-cadherin, or said target is an antibody cocktail specifically binding a combination thereof;   d) a colorectal cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to EpCAM, CD66c, CD66e, CEA, TACSTD2, CK20, CD104, MUC-1, CD318, or N-cadherin, or said target is an antibody cocktail specifically binding a combination thereof;   e) a lung cancer cell and said target that binds said actionable antigen is an antibody that specifically binds to CK18, CK19, TACSTD2, CD318, CD104, CEA, EGFR, or EpCAM, or said target is an antibody cocktail specifically binding a combination thereof;   f) a pancreatic cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to MUC-1, CEA, TACSTD2, CD104, CD318, N-cadherin, MUC-1, or EpCAM, or said target is an antibody cocktail specifically binding a combination thereof;   g) an ovarian cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to MUC-1, TACSTD2, CEA, CD318, CD104, N-cadherin, or EpCAM, or said target is an antibody cocktail specifically binding a combination thereof;   h) an endothelial bladder cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to CD34, CD146, CD62, CD105, CD106, VEGF receptor, or MUC-1, or said target is an antibody cocktail specifically binding a combination thereof;   i) an epithelial bladder cancer cell and said target that binds the actionable antigen is an antibody that specifically binds to TACSTD2, EpCAM, CD318, EGFR, 6B5, N-cadherin or folate binding receptor, or said target is an antibody cocktail that specifically binds a combination thereof;   j) a cancer stem cell and said target that binds the actionable antigen is an antibody that specifically binds to CD133, CD135, CD117, or CD34, or said target is an antibody cocktail that specifically binds a combination thereof;   k) a circulating cancer cell that expresses mesenchymal antigens and said target that binds the actionable antigen is an antibody that specifically binds to FGFR1, FGFR4, EGFR, folate binding receptor, N-cadherin or MSC, or said target is an antibody that specifically binds a combination thereof; and/or   l) a circulating cancer cell that expresses angiogenesis surface antigens and said target that binds the actionable antigen is an antibody that specifically binds to a VEGF receptor.