Method for the production of antibodies

Abstract

The current invention is related to a method for the production of a human monoclonal antibody from a immundeficient non-human animal, said method comprising contacting a new borne immunodeficient non-human animal with a human fetal liver stem cell (FL cell) to generate an immune transplanted non-human animal (reconstituted animal), subsequently contacting said reconstituted animal with a antigen, collecting from said reconstituted animal a human cell producing human antibody against said antigen, and isolating said antibody from said antibody producing cell.

1 . A method for the production of a human monoclonal antibody from a immundeficient non-human animal, said method comprising contacting a new borne immunodeficient non-human animal with a human fetal liver stem cell (FL cell) to generate an immune transplanted non-human animal (reconstituted animal), subsequently contacting said reconstituted animal with a antigen, collecting from said reconstituted animal a human cell producing human antibody against said antigen, and isolating said antibody from said antibody producing cell. 
     
     
         2 . The method of  claim 1  wherein said immunoreconstituted non-human animal is a rodent. 
     
     
         3 . The method of  claim 2  wherein said reconstituted non-human animal is a mouse or a rat. 
     
     
         4 . The method of  claim 3 , wherein said reconstituted mouse is a Rag2 −/− γc −/− , nude Rag 2 −/− , NOD or SCID beige mouse. 
     
     
         5 . The method of  claim 1 , wherein said FL cell is a human hematopoietic fetal liver stem cell (HFL cell). 
     
     
         6 . The method of  claim 5 , wherein said FL cell is combination of a a HFL cell and a human non hematopoeitic fetal liver stem cell. 
     
     
         7 . The method of  claim 1 , wherein said antibody producing cell is a human B-cell. 
     
     
         8 . The method of  claim 1 , wherein the antigen that is contacted with the reconstituted animal is selected from the group consisting of antigen, antigen fragment, MHC/peptide complex, antigen encoding DNA and/or antigen bearing cell. 
     
     
         9 . The method of  claim 1 , wherein said collected cell is a human CD19 +  or CD22 +  cell. 
     
     
         10 . The method of  claim 1 , wherein the fusion partner cell line is a MFP-2, HK-128, K6H6/B5 or Karpas 707 cell line. 
     
     
         11 . A method for the production of a plurality of human B cells producing a plurality of human antibodies against a antigen comprising a) contacting a new borne immune deficient non-human animal with a FL cell to generate a immuno reconstituted animal, b) subsequently contacting said reconstituted animal with a antigen, and c) collecting said plurality of antibody producing human B cells out of said non-human animal. 
     
     
         12 . A method for the recombinant production of an antibody, characterized in contacting a new borne immune deficient non-human animal with a FL cell, subsequently contacting said non-human animal with a antigen, collecting from said non-human animal a human cell producing human antibody against said antigen, and isolating said antibody, sequencing the variable regions, constructing expression vector(s) encoding heavy and/or light chain at least CDRs, combined with a human constant chain, expressing said vector(s) in appropriate host cell(s) and isolating said antibody (immunoreactive protein) from said host cell(s) or fermentation supernatant. 
     
     
         13 . A method for the recombinant production of an antibody, characterized in contacting a new borne immune deficient non-human animal with a FL cell, subsequently contacting said non-human animal with a antigen, collecting from said non-human animal a human cell producing human antibody against said antigen, and isolating mRNA from said human cell producing human antibody, generating antibody specific cDNA (e.g. by use of Immuneglobulin specific primers), constructing expression vector(s) encoding heavy and/or light chain at least CDRs, combined with a human constant chain, expressing said vector(s) in appropriate host cell(s) and isolating said antibody (immunoreactive protein) from said host cell(s) or fermentation supernatant. 
     
     
         14 . A method for the selection of an immortal cell producing a human antibody showing specific binding to a antigen, said method being characterized in providing a plurality of human B cells from the spleen of a non human animal producing a complete plurality of human antibodies, fusing said cells or a subset thereof with an immoral myeloma cell or immortalizing said B cells or subset by EBV transformation, and selecting a hybridoma cell producing a human antibody showing specific binding to said antigen. 
     
     
         15 . A method for the selection of an immortal cell producing an antibody showing specific binding affinity of at least 10 −6  mol/l to a antigen, said method being characterized in providing a plurality of human B cells from the spleen of a non human animal producing a complete plurality of human antibodies, fusing said cells or a subset thereof with an immoral myeloma cell or immortalizing said B cells or subset by EBV transformation, and selecting a hybridoma cell producing an antibody showing specific binding affinity of at least 10 −6  mol/l to an antigen.