Development of pilot scale production process and characterization of a recombinant multiepitope malarial vaccine candidate FALVAC-1A expressed in Escherichia coli
DOI: 10.1016/j.pep.2008.05.018
Title: Development of pilot scale production process and characterization of a recombinant multiepitope malarial vaccine candidate FALVAC-1A expressed in Escherichia coli
Journal Title: Protein Expression and Purification
Volume: 61
Issue: 1
Publication Date: September 2008
Start Page: 57
End Page: 64
Published online: online 7 June 2008
ISSN: 1046-5928
Affiliations:

  • a Bharat Biotech International Limited, Genome Valley, Shameerpet Mandal, Turkapally, R.R. District, Hyderabad 500 078, Andhra Pradesh, India

  • b School of Biotechnology, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad 500 072, Andhra Pradesh, India
  • Abstract: e four human malarial species, Plasmodium falciparum causes most of the mortality associated with malaria. Several approaches are being pursued to develop a suitable malaria vaccine since it may be the most effective weapon to fight against malaria. A highly immunogenic, synthetic protein consisting of 21 epitopes from pre-erythrocytic and blood stages of P. falciparum (FALVAC-1A) was constructed and expressed in Escherichia coli. This vaccine candidate was highly immunogenic and induced protective antibodies in rabbits when produced through lab-scale processes in milligram quantities. In order to take this vaccine candidate for further clinical trial, we optimized the process for industrial scale production and purification. Here we describe various methods used in pilot scale production and characterization of FALVAC-1A. A fed-batch cultivation process in a bioreactor at 10-L scale was optimized to express the protein in high yields as inclusion bodies in E. coli cells with the recombinant plasmids. Methods to solubilize, capture and purify the target protein from the inclusion bodies were optimized and the resultant protein was >95% pure based on SDS–PAGE and RP-HPLC. This protein was then refolded and nativity was confirmed by Far-UV CD spectroscopy. Final purified protein was characterized to estimate yield, purity, mass and confirmed to be free of host cell proteins, nucleic acids and bacterial endotoxins. This study confirms that industrial scale clinical grade FALVAC-1A can be produced in a cost-effective manner for clinical trials.
    Received: 13 March 2008
    Revised: 30 May 2008
    Email: gravi71@gmail.com productionrd02@bharatbiotech.com

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