Growth and Purification of Murine Monoclonal Antibodies

Abstract

Once a hybridoma line has been selected and cloned, it can be expanded and seed stocks cryopreserved for future use. Relatively large amounts of purified MAb may also be required. There are a variety of procedures for this that ensure the establishment of a stable cell line secreting high levels of specific immu-noglobulin. High concentrations of antibody can be generated by growing the line in the peritoneal cavity of mice/rats of the same strain as the tumor cell line donor and spleen cell donor. Antibody is secreted into the ascitic fluid formed within the cavity at a concentration up to 10 mg/mL. However, the ascites will contain immunoglobulins derived from the recipient animal that can be removed by affinity chromatography if desired. Several in vitro culture methods using hollow fibres or dialysis tubing (1) have been developed and are commercially available; this avoids the use of recipient mice/rats and contamination by host immunoglobulins, although contamination with culture medium-derived proteins may be a problem.