Assay for identifying antibody producing cells
The present invention provides a homogeneous assay for identifying an antibody producing cell producing an antibody which binds to a selected antigen comprising: a) providing a population of antibody producing cells; b) incubating said population of antibody producing cells with a selected antigen and a labeled anti-antibody antibody, wherein said anti-antibody antibody is capable of distinguishing cells producing an antibody which binds to the selected antigen from those cells which do not; and c) identifying an antibody producing cell capable of producing an antibody which binds to the selected antigen.
1. A homogeneous assay for identifying an antibody producing cell that produces an antibody that binds to a selected antigen comprising:
providing a population of antibody producing cells;
incubating on a microscope slide the population of antibody producing cells with an antigen, said antigen being coupled to an erythrocyte or a particle, or said antigen being expressed on the surface of a cell, and a labeled anti-antibody antibody wherein the anti-antibody antibody distinguishes antibody producing cells that produce an antibody that binds to the selected antigen from antibody producing cells that do not and wherein the anti-antibody antibody binds to all antibodies produced by the antibody producing cells, regardless of their binding specificity; and
without performing any wash steps, identifying an antibody producing cell that produces an antibody that binds to the selected antigen by detecting the increased concentration of labeled anti-antibody antibodies surrounding said antibody producing cell.
2. The assay of claim 1 where the antigen is coupled to an erythrocyte.
3. The assay of claim 1 where the antigen is coupled to a particle.
4. The assay of claim 3 where the antigen is coupled to a particle via a polyclonal antibody.
5. The assay of claim 4 where the polyclonal antibody is an antibody fragment.
6. The assay of claim 5 where the polyclonal antibody is an antibody Fab, Fab′ or F(ab′) 2 fragment.
7. The assay of claim 1 where the antigen is expressed on the surface of a cell.
8. The assay of claim 7 where the cell is a transfected cell.
9. The assay of claim 7 where the cell is a tumor cell.
10. The assay of claim 1 where the antigen is an infectious agent.
11. The assay of claim 1 where the labeled anti-antibody antibody is labeled is an anti-Fc antibody.
12. The assay of claim 1 where the labeled anti-antibody antibody is labeled with a fluorescent label.
13. The assay of claim 12 where the fluorescently-labeled anti-antibody antibody is labeled with FITC.
14. The assay of claim 13 where the FITC labeled anti-antibody antibody is an anti-Fc antibody.
15. The assay of claim 1 where the antibody producing cells are B cells, plasma cells, plasmablasts, activated B cells or memory B cells.
16. The assay of claim 1 where the antibody producing cell that produces an antibody is a hybridoma cell or a mammalian cell engineered to express antibodies.
17. A method for synthesizing an antibody that binds to a selected antigen comprising:
providing a population of antibody producing cells;
incubating on a microscope slide the population of antibody producing cells with an antigen, said antigen being coupled to an erythrocyte or a particle, or said antigen being expressed on the surface of a cell, and a labeled anti-antibody antibody, wherein the anti-antibody antibody distinguishes antibody producing cells that produce an antibody that binds to the selected antigen from antibody producing cells that do not and wherein the anti-antibody antibody binds to all antibodies produced by the antibody producing cells, regardless of their binding specificity;
without performing any wash steps, identifying an antibody producing cell that produces an antibody that binds to the selected antigen by detecting the increased concentration of labeled anti-antibody antibodies surrounding said antibody producing cell;
isolating the identified antibody producing cell that produces said antibody that binds to the selected antigen; and
directly or indirectly synthesizing said antibody that binds to the selected antigen from the isolated antibody producing cell that produces said antibody.