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

被引：76

作者：

Goud, GN

Bottazzi, ME

Zhan, B

Mendez, S

Deumic, V

Plieskatt, J

Liu, S

Wang, Y

Bueno, L

Fujiwara, R

Samuel, A

Ahn, SY

Solanki, M

Asojo, OA

Wang, J

Bethony, JM

Loukas, A

Roy, M

Hotez, PJ

机构：

[1] George Washington Univ, Med Ctr, Dept Microbiol & Trop Med, Washington, DC 20037 USA

[2] NIAID, Malaria Vaccine Dev Branch, NIH, Bethesda, MD 20892 USA

[3] Univ Nebraska, Med Ctr, Eppley Inst Res Canc & Allied Dis, Omaha, NE 68198 USA

[4] Queensland Inst Med Res, Div Infect Dis & Immunol, Brisbane, Qld 4006, Australia

[5] Sci Applicat Int Corp, Frederick, MD USA

来源：

VACCINE | 2005年 / 23卷 / 39期

关键词：

hookworm; vaccine; Necator americanus; Na-ASP-2;

D O I：

10.1016/j.vaccine.2005.04.040

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

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 (R) and injected into rats to determine immunological potency, three 50 mu g 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. (c) 2005 Elsevier Ltd. All rights reserved.

引用

页码：4754 / 4764

页数：11

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