BIOMAT Database Logo BIOMAT Database Logo

Expression, purification, and molecular analysis of the Necator americanus glutathione S-transferase 1 (Na-GST-1): a production process developed for a lead candidate recombinant hookworm vaccine antigen. 2012

Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Department of Microbiology, Immunology & Tropical Medicine, George Washington University Medical Center, Washington, DC, USA.

The enzyme Necator americanus glutathione S-transferase 1 (Na-GST-1) belongs to a unique Nu class of GSTs and is a lead candidate antigen in a bivalent human hookworm vaccine. Here we describe the expression of Na-GST-1 in the yeast Pichia pastoris at the 20 L manufacturing scale and its purification process performed by three chromatographic steps, comprised of a Q Sepharose XL anion exchange column, followed by a Butyl Sepharose HP hydrophobic affinity column and a Superdex 75 size-exclusion column. Approximately 1.5 g of recombinant protein was recovered at an overall process yield of 51%, with a purity grade of 98% and the absence of detectable host cell protein. By mass spectrometry the recombinant protein exhibits a mass of 23,676Da, which closely matches the predicted molecular mass of the protein. The expression and purification methods described here are suitable for further scale-up product development and for its use to design formulation processes suitable to generate a vaccine for clinical testing.

UI MeSH Term Description Entries
D010843 Pichia Yeast-like ascomycetous fungi of the family Saccharomycetaceae, order SACCHAROMYCETALES isolated from exuded tree sap. Hansenula,Hansenulas,Pichias
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002850 Chromatography, Gel Chromatography on non-ionic gels without regard to the mechanism of solute discrimination. Chromatography, Exclusion,Chromatography, Gel Permeation,Chromatography, Molecular Sieve,Gel Filtration,Gel Filtration Chromatography,Chromatography, Size Exclusion,Exclusion Chromatography,Gel Chromatography,Gel Permeation Chromatography,Molecular Sieve Chromatography,Chromatography, Gel Filtration,Exclusion Chromatography, Size,Filtration Chromatography, Gel,Filtration, Gel,Sieve Chromatography, Molecular,Size Exclusion Chromatography
D002851 Chromatography, High Pressure Liquid Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed. Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D004591 Electrophoresis, Polyacrylamide Gel Electrophoresis in which a polyacrylamide gel is used as the diffusion medium. Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D005982 Glutathione Transferase A transferase that catalyzes the addition of aliphatic, aromatic, or heterocyclic FREE RADICALS as well as EPOXIDES and arene oxides to GLUTATHIONE. Addition takes place at the SULFUR. It also catalyzes the reduction of polyol nitrate by glutathione to polyol and nitrite. Glutathione S-Alkyltransferase,Glutathione S-Aryltransferase,Glutathione S-Epoxidetransferase,Ligandins,S-Hydroxyalkyl Glutathione Lyase,Glutathione Organic Nitrate Ester Reductase,Glutathione S-Transferase,Glutathione S-Transferase 3,Glutathione S-Transferase A,Glutathione S-Transferase B,Glutathione S-Transferase C,Glutathione S-Transferase III,Glutathione S-Transferase P,Glutathione Transferase E,Glutathione Transferase mu,Glutathione Transferases,Heme Transfer Protein,Ligandin,Yb-Glutathione-S-Transferase,Glutathione Lyase, S-Hydroxyalkyl,Glutathione S Alkyltransferase,Glutathione S Aryltransferase,Glutathione S Epoxidetransferase,Glutathione S Transferase,Glutathione S Transferase 3,Glutathione S Transferase A,Glutathione S Transferase B,Glutathione S Transferase C,Glutathione S Transferase III,Glutathione S Transferase P,Lyase, S-Hydroxyalkyl Glutathione,P, Glutathione S-Transferase,Protein, Heme Transfer,S Hydroxyalkyl Glutathione Lyase,S-Alkyltransferase, Glutathione,S-Aryltransferase, Glutathione,S-Epoxidetransferase, Glutathione,S-Transferase 3, Glutathione,S-Transferase A, Glutathione,S-Transferase B, Glutathione,S-Transferase C, Glutathione,S-Transferase III, Glutathione,S-Transferase P, Glutathione,S-Transferase, Glutathione,Transfer Protein, Heme,Transferase E, Glutathione,Transferase mu, Glutathione,Transferase, Glutathione,Transferases, Glutathione
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000947 Antigens, Helminth Any part or derivative of a helminth that elicits an immune reaction. The most commonly seen helminth antigens are those of the schistosomes. Helminth Antigens
D015801 Helminth Proteins Proteins found in any species of helminth. Helminth Protein,Protein, Helminth,Proteins, Helminth
D017221 Necator americanus A common parasite of humans in the moist tropics and subtropics. These organisms attach to villi in the small intestine and suck blood causing diarrhea, anorexia, and anemia. Hookworm, New World,Hookworms, New World,New World Hookworm,New World Hookworms

Related Publications

Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Optimization and revision of the production process of the Necator americanus glutathione S-transferase 1 (Na-GST-1), the lead hookworm vaccine recombinant protein candidate.
January 2014, Human vaccines & immunotherapeutics,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Structure of glutathione S-transferase 1 from the major human hookworm parasite Necator americanus (Na-GST-1) in complex with glutathione.
September 2014, Acta crystallographica. Section F, Structural biology communications,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Glutathione S-transferase (GST) expression in the human hookworm Necator americanus: potential roles for excretory-secretory forms of GST.
June 1995, Acta tropica,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Structure of monomeric Na-GST-3, a glutathione S-transferase from the major human hookworm parasite Necator americanus.
August 2013, Acta crystallographica. Section F, Structural biology and crystallization communications,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Expression, purification, and characterization of the Necator americanus aspartic protease-1 (Na-APR-1 (M74)) antigen, a component of the bivalent human hookworm vaccine.
January 2015, Human vaccines & immunotherapeutics,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Antigen expression during development of the human hookworm, Necator americanus (Nematoda).
March 1987, Parasite immunology,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Potency testing for the experimental Na-GST-1 hookworm vaccine.
October 2010, Expert review of vaccines,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Computer-aided identification of potential inhibitors against Necator americanus glutathione S-transferase 3.
January 2022, Informatics in medicine unlocked,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Production and characterization of a monoclonal antibody against recombinant glutathione S-transferase (GST) of Fasciola gigantica.
December 2002, Asian Pacific journal of allergy and immunology,
Gaddam Narsa Goud, and Vehid Deumic, and Richi Gupta, and Jill Brelsford, and Bin Zhan, and Portia Gillespie, and Jordan L Plieskatt, and Eric I Tsao, and Peter J Hotez, and Maria Elena Bottazzi
Analysis of the glutathione S-transferase (GST) gene family.
November 2004, Human genomics,
Copied contents to your clipboard!