We utilized the silkworm (Bombyx mori)/baculovirus system to produce recombinant Major Merozoite Surface Protein (1) (MSP142) because of the low cost and potential high yield of this expression system. The MSP142 (3D7 sequence) was cloned into the baculovirus, BmNPV with the melittin signal sequence. The recombinant virus, BmNPV-Sp42 was used to infect silkworms for the expression of MSP142 (Sp42). One recombinant clone expressed high level of Sp42 with an estimated 0.5 mg of antigen produced within a single worm. The Sp42 was recognized by monoclonal and polyclonal antibodies specific for parasite MSP1 in direct binding and competitive binding ELISAs, suggesting that Sp42 possesses antigenic determinants similar to parasite MSP142. Immunogenicity studies were performed in rabbits. Sp42 induced high titers of antibodies crossreactive with MSP1. Specificity analyses showed that anti-Sp42 antibodies reacted primarily against conserved determinants on MSP1-19. Our results showed that the silkworm expression system can produce recombinant MSP142 that are antigenically and immunogenically comparable to other recombinant MSP1 antigens expressed in other eukaryotic systems. The low cost at high level of protein expression makes it an attractive alternative for the development of a human malaria vaccine.
申请公布号
WO0134188(A1)
申请公布日期
2001.05.17
申请号
WO2000US31064
申请日期
2000.11.09
申请人
UNIVERSITY OF HAWAII;THE CHINESE UNIVERSITY OF HONG KONG;THE QUEEN EMMA FOUNDATION;HUI, GEORGE, S., N.;LAP-YIN, PANG;HO, WALTER, K., K.
发明人
HUI, GEORGE, S., N.;LAP-YIN, PANG;HO, WALTER, K., K.