Genetic engineering of an H-2Dd/Q10b chimeric histocompatibility antigen: purification of soluble protein from transformant cell supernatants.
We have constructed a recombinant class I gene in which 5' sequences of H-2Dd 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-2Dd 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-2Dd monoclonal antibodies that identify epitopes on the N and C1 domains as well as with an anti-Q10 carboxyl-terminal peptide antiserum. The H-2Dd-derived antigen is associated with beta 2-microglobulin and is readily purified in milligram amounts from culture supernatants by immunoaffinity chromatography.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D006183:H-2 Antigens / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D064532:Histocompatibility Antigen H-2D; D007739:L Cells / Q000032:analysis; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics* / Q000302:isolation & purification; D014170:Transformation, Genetic
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006183:H-2 Antigens; D064532:Histocompatibility Antigen H-2D; D011994:Recombinant Proteins