A Method for Monoclonal Antibody Isotype Switching: Anti-CD60 V H Expression in a Heavy Chain-Deficient Hybridoma Variant

Abstract

Alteration of monoclonal antibody isotype is desirable for a variety of purposes, including obtaining an improved reagent for investigative or therapeutic use. A variety of approaches for isotype switching, particularly from IgM to various IgG subclasses, have been described. Antibodies that recognize carbohydrate determinants on glycoproteins, glycolipids, or polysaccharides are generally of the IgM isotype. This includes all available antibodies to the human CD60 antigen, a determinant with cell coactivating properties described on a subset of T lymphocytes and on other cell types. In this report a new method for monoclonal antibody isotype switching is presented. A plasmid containing the V_H regions of anti-CD60 linked to C_γ1 was transfected into a spontaneously arising variant of the CD60 hybridoma that produced κ light chain but no heavy chain. This transfected hybridoma line maintains stable production of useful quantities of IgG₁ monoclonal anti-CD60 in vitro and in vivo.