Protein-A antibody-binding TPAAB) technique for detection of tumor-associated membrane antigen (TAMA) of Lewis lung carcinoma.

Xenoantisera were prepared in rabbits against intact paraformaldehyde-fixed Lewis lung carcinoma (LLCa) cells and membrane solubilized proteins extracted with Triton X-100 and 3M KCI. The antisera were analyzed for antibodies to tumor-associated membrane antigens (TAMA) by means of a solid-phase radioimmunoassay (RIA). THe immunoassay employed 125-labelled Staphylococcus aureus protein A (SPA) to detect specific antibody binding to glutaraldehyde-fixed Lewis lung tumor target cells. The protein-A antibody-binding (PAAB) method is rapid, sensitive and reproducible. The PAAB revealed a possible LLCa-TAMA as evidenced by the reactivity of xenoantisera raised to LL paraformaldehyde tumor cell vaccine (TCV) and LL tumor Triton X-100 (TTri) and 3M KCI (TKCI) extracts of membrane preparations. The reactivity of our antisera followed linear dose-response kinetics but the degree of reactivity decreased from anti-TCV to anti-TTri to anti-TKCI respectively. The LL-TAMA demonstrated specificity since our antisera did not significantly cross-react with normal C56BL/6 cellular antigens or other known murine tumor-associated surface antigens of virally and chemically-induced tumor systems.