Methods for Isolating Immune Binding Proteins

Abstract

Described herein are immune binding proteins and method for obtaining immune binding proteins from genomic or other sources. Also described herein are nucleic acids encoding the immune binding proteins in which the natural multimeric association of chains is maintained in the nucleic acids and the immune binding proteins made therefrom. For example, nucleic acids encoding antibodies that are amplified from a B-cell using the methods described herein maintain the natural pairing of heavy and light chains from the B-cell. This maintenance of pairing (or multimerization) produces libraries and/or repertoires of immune binding proteins that are enriched for useful binding molecules.

1 . (canceled) 
     
     
         2 . (canceled) 
     
     
         3 . (canceled) 
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . (canceled) 
     
     
         7 . (canceled) 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . (canceled) 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . (canceled) 
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . (canceled) 
     
     
         21 . A method of isolating a target cell of interest, wherein the target cell comprises an antibody encoded by a plurality of nucleic acids, comprising the steps of: mixing a repertoire of bait particles with a repertoire of target cells, wherein repertoire of bait particles comprises a repertoire of antigens, wherein a bait particle in the repertoire of bait particles comprises an antigen of interest and an oligonucleotide, wherein the oligonucleotide comprises a barcode that is specific for the antigen of interest; isolating a bait particle with the target cell of interest bound to the antigen of interest; sequencing the barcode from the isolated bait particle with the target cell of interest to identify the antigen of interest; lysing the target cell; and sequencing the plurality of nucleic acids that encode the antibody in the target cell of interest, whereby a target cell of interest is isolated and identified. 
     
     
         22 . The method of  claim 21 , wherein the bait particle is a magnetic bead. 
     
     
         23 . The method of  claim 21 , wherein the bait particle is a bead with a fluorophore. 
     
     
         24 . The method of  claim 21 , wherein the target cell is a B-cell with an antibody on a cell surface. 
     
     
         25 . The method of  claim 21 , wherein the bait particle further comprises a second oligonucleotide; wherein the second oligonucleotide comprises a bait particle specific barcode sequence. 
     
     
         26 . The method of  claim 21 , wherein the bait particle further comprises a second oligonucleotide; wherein the second oligonucleotide binds to a nucleic acid in the target cell. 
     
     
         27 . The method of  claim 21 , wherein the immune binding protein is displayed on a surface of the target cell using a display technology, and wherein the step of obtaining sequence information from the target cell of interest comprises sequencing a nucleic acid that encodes the antibody displayed on the surface of the target cell of interest. 
     
     
         28 . The method of  claim 27 , wherein the display technology is a cell display technology. 
     
     
         29 . The method of  claim 27 , wherein an antibody variable domain is displayed by the display technology. 
     
     
         30 . The method of  claim 21 , wherein the antigen of interest is from an influenza virus. 
     
     
         31 . The method of  claim 30 , wherein the antigen of interest is a haemagluttinin from an isolate of influenza. 
     
     
         32 . The method of  claim 31 , wherein the repertoire of bait particles has another bait particle with a different influenza antigen, wherein the other bait particle has an oligonucleotide that includes a different bar code. 
     
     
         33 . The method of  claim 32 , wherein the different influenza antigen is a haemagluttinin from an different isolate of influenza. 
     
     
         34 . The method of  claim 21 , wherein the bait particle further comprises another oligonucleotide, wherein the other oligonucleotide comprises a primer for the plurality of nucleic acids that encode the antibody in the target cell of interest. 
     
     
         35 . The method of  claim 21 , wherein multiple bait particles from the repertoire of bait particles are attached to the target cell. 
     
     
         36 . The method of  claim 21 , wherein the repertoire of target cells are obtained from an individual that had an immune response to the antigen of interest.