Genetic engineering of an H-2D(d)/Q10b chimeric histocompatibility antigen: Purification of soluble protein from transformant cell supernatants

Abstract

We have constructed a recombinant class I gene in in which 5' sequences of H-2D(d) are linked to the 3' half of a Qa subregion gene, Q10b. This hybrid gene would be expected to direct the synthesis of a protein containing the N and C1 domains of H-2D(d) covalently linked to the C2 domain of the secreted, nonpolymorphic, Q10b antigen. Following DNA-mediated gene transfer into mouse L cells, transformants were analyzed by radiolabeling and immunoprecipitation. These cells secreted a molecule reactive with anti-H-2D(d) monoclonal antibodies that identify epitopes on the N and C1 domains as well as with an anti-Q10 carboxyl-terminal peptide antiserum. The H-2D(d)-derived antigen is associated wi..