Production and single-step purification of Brugia malayi abundant larval transcript (ALT-2) using hydrophobic interaction chromatography.
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).
Bhuvanesh
Surianarayanan
S
Centre for Biotechnology, Anna University, Chennai 600025, India.
Arunkumar
Chakkaravarthi
C
Kaliraj
Perumal
P
Ramalingam
Subramanian
S
D000947:Antigens, Helminth / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D001709:Biotechnology / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D057927:Hydrophobic and Hydrophilic Interactions; D007544:Isopropyl Thiogalactoside / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D015533:Transcriptional Activation; D015854:Up-Regulation
C521555:ALT-2 protein, Brugia malayi; D000947:Antigens, Helminth; D003470:Culture Media; D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside