Journal of Industrial Microbiology & Biotechnology

, Volume 37, Issue 10, pp 1053–1059

Production and single-step purification of Brugia malayi abundant larval transcript (ALT-2) using hydrophobic interaction chromatography

Original Paper

Abstract

Abundant larval transcript (ALT), a novel filarial protein, has been shown to have great potential as a vaccine in the prevention of human lymphatic filariasis. In this study, we report a method for the production of recombinant ALT-2 protein, expressed in the cytoplasm of bacterium Escherichia coli in soluble form and purification in a single step using hydrophobic interaction chromatography (HIC). Fermentation was done by continuous fed-batch methodology with dissolved oxygen (DO)-controlled feed addition. The culture was induced with 1 mM isopropyl-β-d-thiogalactopyranoside (IPTG). Up to 9 g/l dry cell weight (DCW) of biomass was obtained from 1.6 l of Luria–Bertani (LB) broth in a bench-scale reactor. Around 200 mg/l of purified ALT-2 with a yield of about 60% was obtained. This is almost a 2.5-fold increase in final protein yield compared to purification using immobilized metal affinity chromatography (IMAC).

Keywords

Recombinant vaccine antigen Lymphatic filariasis Abundant larval transcript (ALT-2) Fed-batch DO-stat Hydrophobic interaction chromatography (HIC) 

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Copyright information

© Society for Industrial Microbiology 2010

Authors and Affiliations

  1. 1.Centre for BiotechnologyAnna UniversityChennaiIndia

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