Bioprocess and Biosystems Engineering

, Volume 33, Issue 6, pp 719–730

Identification of cultivation condition to produce correctly folded form of a malaria vaccine based on Plasmodium falciparum merozoite surface protein-1 in Escherichia coli

Original Paper

DOI: 10.1007/s00449-009-0394-x

Cite this article as:
Mazumdar, S., Sachdeva, S., Chauhan, V.S. et al. Bioprocess Biosyst Eng (2010) 33: 719. doi:10.1007/s00449-009-0394-x

Abstract

The C-terminal, 19-kDa domain of Plasmodium falciparum merozoite surface protein-1 (PfMSP-119) is among the leading vaccine candidate for malaria due to its essential role in erythrocyte invasion by the parasite. We designed a synthetic gene for optimal expression of recombinant PfMSP-119 in Escherichia coli and developed a scalable process to obtain high-quality PfMSP-119. The synthetic gene construct yielded a fourfold higher expression level of PfMSP-119 in comparison to the native gene construct. Optimization of cultivation conditions in the bioreactor indicated important role of yeast extract and substrate feeding strategy for obtaining enhanced expression of soluble and correctly folded PfMSP-119. It was observed that the higher expression level of PfMSP-119 was essentially associated with the generation of higher level of incorrectly folded PfMSP-119. A simple purification procedure comprising metal affinity and ion exchange chromatography was developed to purify correctly folded form of PfMSP-119 from cell lysate. Biochemical and biophysical characterization of purified PfMSP-119 suggested that it was highly pure, homogeneous, and correctly folded.

Keywords

Malaria vaccine Merozoite surface protein-1 Recombinant Escherichia coli Process development Characterization 

Supplementary material

449_2009_394_MOESM1_ESM.pdf (23 kb)
Supplementary Figures (PDF 23 kb)

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Malaria Research GroupInternational Centre for Genetic Engineering and Biotechnology (ICGEB)New DelhiIndia

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