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Vaccine. 2005 Sep 15;23(39):4754-64.

Expression of the Necator americanus hookworm larval antigen Na-ASP-2 in Pichia pastoris and purification of the recombinant protein for use in human clinical trials.

Author information

1
Department of Microbiology and Tropical Medicine, The George Washington University Medical Center, Ross Hall 736, 2300 Eye Street NW, Washington, DC 20037, USA. mtmgng@gwumc.edu

Abstract

The ASP-2 protein secreted by infective larvae of the human hookworm, Necator americanus, is under development as a recombinant vaccine. Recombinant Na-ASP-2 was expressed in Pichia pastoris, and the purified protein was characterized. At the 60 L scale, the 21.3 kDa recombinant protein was produced at a yield of 0.4 g/L. When formulated with Alhydrogel and injected into rats to determine immunological potency, three 50 microg doses of the formulated recombinant protein elicited geometric mean antibody titers up to 1:234,881. Rat anti-Na-ASP-2 antibody recognized larval-derived ASP-2 and also inhibited larval migration through skin in vitro. The processes developed and tested for the high yield production of recombinant Na-ASP-2 provide a foundation for clinical vaccine development.

PMID:
16054275
DOI:
10.1016/j.vaccine.2005.04.040
[Indexed for MEDLINE]

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