Radioimmunoassay for detecting antibodies against murine malarial parasite antigens: monoclonal antibodies recognizing Plasmodium yoelii antigens.

A solid-phase radioimmunoassay (SPRIA) in microtiter wells was established for detecting antibodies against Plasmodium yoelii Ag. The SPRIA was found 1) to require as little as 5 micrograms of crude parasite Ag per well, 2) to be able to detect 0.5 ng of monoclonal Ab, and 3) to be 10(4) times more sensitive than the indirect fluorescent Ab staining technique. In a modification of the above assay using intact RBC as an Ag, hyperimmune serum showed significant binding to the surface of erythrocytes of mice infected with P. yoelii parasites but not to RBC of normal mice. Hybridomas were prepared by fusing infected mouse spleen cells with P3-NS1/I-Ag4-1 myeloma cells. Using the SPRIA, hybrids secreting Ab against P. yoelii 17XL Ag were detected. All the hybrids detected by SPRIA secreted Ab binding to free parasites but not to the surface membrane Ag on the infected RBC. The number of positive hybrids produced by fusing spleen cells from mice given a double infection (17XNL and 17 XL) was not greater than that produced by freezing spleen cells from mice given a single infection (17 XL).