Production of a monoclonal antibody to the arylphorin of Heliothis zea

Abstract

AbstractA species‐specific monoclonal antibody was produced to whole plasma of fifth instar larvae of the corn earworm, Heliothis zea (Boddie) (Lepidoptera:Noctuidae). This antibody did not cross‐react with proteins from the plasma of any of the other lepidopteran larvae tested, including other Heliothis spp. The antigen recognized by this antibody was characterized and found to be arylphorin, a prominent larval storage protein. This conclusion was based on the electrophoretic as well as immunological characteristics of the antigen. Polyacrylamide gel electrophoresis indicated that the antigen had an apparent native molecular weight of 460,000, and that it was composed of subunits having apparent molecular weights of 76,000. The isoelectric point of the antigen was 5.9, with some microheterogeneity being seen. Western blotting of arylphorin against the monoclonal antibody clearly identified the antigen as arylphorin. This protein was not found in egg homogenates or early instar larval plasma, but it was present in large quantities in pupal homogenates and, at trace levels, in adult homogenates. The efficient production and selection of hybridomas producing antibodies specific to H. zea arylphorin using unfractionated plasma as immunogen illustrates the fact that monoclonal antibody technology can produce highly specific antibodies using crude antigen preparations. We discuss the tradeoffs one must accept when choosing this strategy over one using purified immunogens.