DEVELOPMENT AND PARTIAL CHARACTERIZATION OF MONOCLONAL-ANTIBODIES TO VENOM OF THE PARASITOID MICROPLITIS-DEMOLITOR

The venom of Microplitis demolitor consists of a mixture of proteins. On native PAGE gels three major proteins designated a, b, and g were d etected, while on SDS-PAGE gels two major proteins of M(r) 64.5 and 30 .8 kD and several minor proteins were detected. No proteins smaller th an M(r) 30.8 kD were present. Murine monoclonal antibodies were genera ted against different venom components. Analysis by Western blot of ve nom proteins separated on native and SDS-PACE gels confirmed that anti bodies from seven hybridoma lines recognized venom components. Two of the seven hybridoma lines reacted specifically with protein g on nativ e PAGE gels and the M(r) 30.8 k protein on SDS-PACE gels, while four o ther lines cross-reacted with these and other venom proteins. The fina l hybridoma line reacted with protein a when venom was separated on na tive PACE gels and an array of proteins when venom was separated on SD S-PAGE gels. Using an enzyme-immunoassay and specific monoclonal antib odies, M. demolitor females were estimated to inject 0.02-0.05 venom g land reservoir equivalents into its host, Pseudoplusia includens, at o viposition. Venom proteins persisted in host hemolymph for 6-12 h befo re dropping to undetectable levels. (C) 1994 Wiley-Liss, Inc.