title: 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.
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.
authors.affiliation: 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
authors.forename: Gaddam Narsa
authors.initials: GN
authors.lastname: Goud
authors.forename: Maria Elena
authors.initials: ME
authors.lastname: Bottazzi
authors.forename: Bin
authors.initials: B
authors.lastname: Zhan
authors.forename: Susana
authors.initials: S
authors.lastname: Mendez
authors.forename: Vehid
authors.initials: V
authors.lastname: Deumic
authors.forename: Jordan
authors.initials: J
authors.lastname: Plieskatt
authors.forename: Sen
authors.initials: S
authors.lastname: Liu
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Lilian
authors.initials: L
authors.lastname: Bueno
authors.forename: Ricardo
authors.initials: R
authors.lastname: Fujiwara
authors.forename: Andre
authors.initials: A
authors.lastname: Samuel
authors.forename: So Yeong
authors.initials: SY
authors.lastname: Ahn
authors.forename: Maneesha
authors.initials: M
authors.lastname: Solanki
authors.forename: Oluwatoyin A
authors.initials: OA
authors.lastname: Asojo
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Bethony
authors.forename: Alex
authors.initials: A
authors.lastname: Loukas
authors.forename: Michael
authors.initials: M
authors.lastname: Roy
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Hotez
mesh_terms: 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*
chemical_list: D015801:Helminth Proteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic