Journal
PROTEIN EXPRESSION AND PURIFICATION
Volume 83, Issue 2, Pages 145-151Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2012.03.013
Keywords
Hookworm; Vaccine; Vaccines; Sabin Vaccine Institute; Necator americanus; Na-GST-1; Albendazole; Mebendazole
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Funding
- Sabin Vaccine Institute
- Bill & Melinda Gates Foundation [32472, 38988]
- Dutch Ministry of Foreign Affairs
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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 23,676 Da, 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. (C) 2012 Elsevier Inc. All rights reserved.
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