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
GN Goud, ME Bottazzi, B Zhan, S Mendez, V Deumic… - Vaccine, 2005 - Elsevier
GN Goud, ME Bottazzi, B Zhan, S Mendez, V Deumic, J Plieskatt, S Liu, Y Wang, L Bueno…
Vaccine, 2005•ElsevierThe 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 60L 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 50μg doses of
the formulated recombinant protein elicited geometric mean antibody titers up to 1: 234,881 …
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 60L 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 50μg doses of
the formulated recombinant protein elicited geometric mean antibody titers up to 1: 234,881 …
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 60L scale, the 21.3kDa recombinant protein was produced at a yield of 0.4g/L. When formulated with Alhydrogel® and injected into rats to determine immunological potency, three 50μg 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.
Elsevier
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