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Infection and Immunity, August 2002, p. 4471-4476, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4471-4476.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

High-Level Expression of the Malaria Blood-Stage Vaccine Candidate Plasmodium falciparum Apical Membrane Antigen 1 and Induction of Antibodies That Inhibit Erythrocyte Invasion

Clemens H. M. Kocken,1 Chrislaine Withers-Martinez,2,3 Martin A. Dubbeld,1 Annemarie van der Wel,1 Fiona Hackett,2 Michael J. Blackman,2 and Alan W. Thomas1*

Department of Parasitology, Biomedical Primate Research Centre, 2280 GH Rijswijk, The Netherlands,1 Division of Parasitology,2 Division of Protein Structure, National Institute for Medical Research, Mill Hill, London NW7 1AA, United Kingdom3

Received 9 November 2001/ Returned for modification 1 March 2002/ Accepted 13 May 2002

Apical membrane antigen 1 (AMA-1) is a highly promising malaria blood-stage vaccine candidate that has induced protection in rodent and nonhuman primate models of malaria. Authentic conformation of the protein appears to be essential for the induction of parasite-inhibitory antibody responses. Here we have developed a synthetic gene with adapted codon usage to allow expression of Plasmodium falciparum FVO strain AMA-1 (PfAMA-1) in Pichia pastoris. In addition, potential N-glycosylation sites were changed, exploiting the lack of conservation of these sites in Plasmodium, to obtain high-level secretion of a homogeneous product, suitable for scale-up according to current good manufacturing procedures. Purified PfAMA-1 displayed authentic antigenic properties, indicating that the amino acid changes had no deleterious effect on the conformation of the protein. High-titer antibodies, raised in rabbits, reacted strongly with homologous and heterologous P. falciparum by immunofluorescence. In addition, purified immunoglobulin G from immunized animals strongly inhibited invasion of red blood cells by homologous and, to a somewhat lesser extent, heterologous P. falciparum.

* Corresponding author. Mailing address: BPRC, Department of Parasitology, Lange Kleiweg 139, 2288 GJ Rijswijk, The Netherlands. Phone: (31) 15-2842 538. Fax: (31) 15-2843 986. E-mail: thomas{at}bprc.nl.

Editor: J. M. Mansfield

Infection and Immunity, August 2002, p. 4471-4476, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4471-4476.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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