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Infection and Immunity, June 2002, p. 2772-2779, Vol. 70, No. 6
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.6.2772-2779.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

In Vivo Expression and Immunological Studies of the 42-Kilodalton Carboxyl-Terminal Processing Fragment of Plasmodium falciparum Merozoite Surface Protein 1 in the Baculovirus-Silkworm System

Alan L. Y. Pang,1 Caryn N. Hashimoto,2 Leslie Q. Tam,2 Z. Q. Meng,3 George S. N. Hui,2 and Walter K. K. Ho1*

Department of Biochemistry, Chinese University of Hong Kong, Shatin, Hong Kong,1 Department of Tropical Medicine and Medical Microbiology, University of Hawaii, Honolulu, Hawaii 96816,2 Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang, People's Republic of China3

Received 22 August 2001/ Returned for modification 24 October 2001/ Accepted 12 February 2002

The 42-kDa carboxyl-terminal processing fragment of Plasmodium falciparum merozoite surface protein 1 (MSP-142) is an anti-erythrocytic stage malaria vaccine candidate. In this study, MSP-142 was expressed by using the Bombyx mori nuclear polyhedrosis virus-silkworm expression system, and the antigenicity and immmunogenicity of the recombinant protein, Bmp42, were evaluated. The average yield of Bmp42, as determined by a sandwich enzyme-linked immunosorbent assay (ELISA), was 379 µg/ml of infected silkworm hemolymph, which was >100-fold higher than the level attainable in cell culture medium. N-terminal amino acid sequencing revealed that Bmp42 was correctly processed in silkworm cells. Data from immunoblotting, as well as from the inhibition ELISA, suggested that the conformational B-cell epitopes of MSP-142 were recreated in Bmp42. Immunization of rabbits with Bmp42 in complete Freund's adjuvant generated high-titer antibody responses against the immunogen. Specificity analyses of the anti-Bmp42 antibodies using several recombinant MSP-119 proteins expressing variant and conserved B-cell epitopes suggested that the anti-Bmp42 antibodies recognized primarily conserved epitopes on MSP-119. Furthermore, the anti-Bmp42 antibodies were highly effective in inhibiting the in vitro growth of parasites carrying homologous or heterologous MSP-142. Our results demonstrated that the baculovirus-silkworm expression system could be employed to express biologically and immunologically active recombinant MSP-142 at elevated levels; thus, it is an attractive alternative for producing a protective MSP-142 vaccine for human use.

* Corresponding author. Mailing address: Department of Biochemistry, The Chinese University of Hong Kong, Shatin, Hong Kong. Phone: (852) 2609 6345. Fax.: (852) 2603 5123. E-mail: walterk{at}cuhk.edu.hk.

Editor: S. H. E. Kaufmann

Infection and Immunity, June 2002, p. 2772-2779, Vol. 70, No. 6
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.6.2772-2779.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.





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