SYSTEM FOR PRODUCTION OF ANTIBODIES AND THEIR DERIVATIVES

摘要

The present disclosure provides methods and compositions for the production of chimeric antibodies that specifically bind an antigen of interest.

主权项

1. A method for producing a chimeric immunoglobulin-G (IgG) antibody that specifically binds an antigen of interest comprising:
a) isolating nucleic acid sequences encoding IgG heavy and light chain variable regions from a single immune cell producing an IgG that specifically binds the antigen of interest; b) cloning the nucleic acid sequences of part a) into separate expression vectors comprising the IgG heavy or light chain constant regions, or into a single expression vector comprising both the IgG heavy and light chain constant regions; c) introducing the expression vector(s) of part b) into a host cell; d) establishing a stable cell line from the host cell of part c); and e) isolating the IgG produced by the stable cell line of part d), wherein the method comprises simultaneous cloning of the IgG heavy and light chain variable regions isolated from the immune cell of part a), and wherein the expression vector of part b) allows for (i) unidirectional insertion of the IgG heavy and light chain variable regions into the vector, and (i) positive selection of expression vectors comprising cloned sequences.