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.
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.
Goud
Gaddam Narsa
GN
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
Bottazzi
Maria Elena
ME
Zhan
Bin
B
Mendez
Susana
S
Deumic
Vehid
V
Plieskatt
Jordan
J
Liu
Sen
S
Wang
Yan
Y
Bueno
Lilian
L
Fujiwara
Ricardo
R
Samuel
Andre
A
Ahn
So Yeong
SY
Solanki
Maneesha
M
Asojo
Oluwatoyin A
OA
Wang
Jin
J
Bethony
Jeffrey M
JM
Loukas
Alex
A
Roy
Michael
M
Hotez
Peter J
PJ
D000818:Animals; D002986:Clinical Trials as Topic; D015801:Helminth Proteins / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007814:Larva / Q000276:immunology; D017221:Necator americanus / Q000276:immunology*; D010843:Pichia / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000276:immunology*
D015801:Helminth Proteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic