Enhanced Production of Protective Antigen, a Potent Diagnostic Protein of Bacillus anthracis, the Causative Agent of Anthrax

  • Manoj Kumar DRDO-Defence Research and Development Establishment, Gwalior - 474 002, India
  • Nidhi Puranik DRDO-Defence Research and Development Establishment, Gwalior - 474 002, India
  • Nagesh Tripathi DRDO-Defence Research and Development Establishment, Gwalior - 474 002, India
  • Vijai Pal DRDO-Defence Research and Development Establishment, Gwalior - 474 002, India
  • Ajay Goel DRDE, Gwalior
Keywords: Anthrax, Protective antigen, Codon optimization, Ion exchange chromatography, Diagnosis.

Abstract

Protective antigen (PA) produced by Bacillus anthracis is a highly immunogenic protein. Therefore, it has significant importance in serodiagnosis as well as a vaccine candidate for anthrax. In the present study, codons for PA gene were optimised and synthesised for its expression in Escherichia coli. Various expression conditions were optimised for scaled up production of rPA. The final yield of affinity chromatography purified protein was 40.8 mg/l during batch fermentation. For further purification, affinity purified protein was diafiltered and subjected to anion exchange chromatography. SDS-PAGE and Western blot was used to characterise the purified rPA protein. The diagnostic potential of purified rPA was evaluated in Western blot using standards reference serum AVR 801 and cutaneous anthrax clinical sera. The results of the present study established the optimum production of rPA in E. coli after codon optimisation for its subsequent use in diagnosis of anthrax infection.

Author Biographies

Manoj Kumar, DRDO-Defence Research and Development Establishment, Gwalior - 474 002, India

Mr Manoj Kumar received MSc from Jiwaji University, Gwalior, M.P. India, in 2011. Currently working as SRF at DRDO-Defence Research & Development Establishment and pursuing PhD from Bharathiar University, Coimbatore. He is working on identification and characterisation of immunodominant antigen(s): evaluation as subunit vaccine candidate(s) in a mouse model against Bacillus anthracis.

Nidhi Puranik, DRDO-Defence Research and Development Establishment, Gwalior - 474 002, India

Ms Nidhi Puranik received the MSc from Devi Ahilya Vishwavidyalaya, Indore, India in 2009. Currently working as SRF at DRDO-Defence Research & Development Establishment and pursuing PhD from Bharathiar University, Coimbatore. She is working on development of rapid immunological systems for detection and diagnosis of anthrax from environmental and clinical samples.

Nagesh Tripathi, DRDO-Defence Research and Development Establishment, Gwalior - 474 002, India

Dr Nagesh Tripathi received his PhD (Chemical Engineering) from National Institute of Technology, Rourkela. Presently working as Scientist ‘D’ at the DRDO-Defence Research and Development Establishment, Gwalior. His research interest includes : Scale up of biomolecules including recombinant proteins, activated carbon spheres and development of chemical protective suit.

Vijai Pal, DRDO-Defence Research and Development Establishment, Gwalior - 474 002, India

Dr Vijai Pal did his MSc (Biotechnology) from CCS Haryana Agricultural University, Hisar, in 2000 and PhD from Jiwaji University, Gwalior, in 2016. Presently working as Scientist ‘E’ at Defence Research and Development Establishment, Gwalior on development of diagnostic/detection systems for biothreat agents. He has published more than 30 research paper in Journals, besides one book and has filed one Indian Patent.

Published
2019-10-21
How to Cite
Kumar, M., Puranik, N., Tripathi, N., Pal, V., & Goel, A. (2019). Enhanced Production of Protective Antigen, a Potent Diagnostic Protein of Bacillus anthracis, the Causative Agent of Anthrax. Defence Life Science Journal, 4(4), 250-255. https://doi.org/10.14429/dlsj.4.15132
Section
Special Issue Paper