A novel approach for preparing anti-T cell receptor constant region antibodies

To obtain antibodies against the individual chains of the T cell receptor (TCR) complex, we have produced chimeric proteins containing domains from immunoglobulin (Ig) and TCR polypeptide chains. Basically, the Ig light chains were used as carriers for the TCR constant (C) region domains. The exons which encode the main body of the C regions of the alpha, beta and the related gamma polypeptide chains were "engineered" into the intronic region between the rearranged Ig variable (V) region and C kappa region genes. All three chimeric genes were expressed in myeloma cells, and the proteins of expected apparent molecular weight were produced. Secreted proteins containing the C beta domain were purified from the culture supernatant by using anti-kappa antibody affinity columns, and two rabbits were then immunized with the purified protein. Both rabbits produced antibodies able to immunoprecipitate the heterodimeric TCR protein.