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Yield Optimisation of Hepatitis B Virus Core Particles in E. Coli Expression System for Drug Delivery Applications

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Journal Sci Rep
Specialty Science
Date 2017 Mar 4
PMID 28256592
Citations 8
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Abstract

An E. coli expression system offers a mean for rapid, high yield and economical production of Hepatitis B Virus core (HBc) particles. However, high-level production of HBc particles in bacteria is demanding and optimisation of HBc particle yield from E. coli is required to improve laboratory-scale productivity for further drug delivery applications. Production steps involve bacterial culture, protein isolation, denaturation, purification and finally protein assembly. In this study, we describe a modified E. coli based method for purifying HBc particles and compare the results with those obtained using a conventional purification method. HBc particle morphology was confirmed by Atomic Force Microscopy (AFM). Protein specificity and secondary structure were confirmed by Western Blot and Circular Dichroism (CD), respectively. The modified method produced ~3-fold higher yield and greater purity of wild type HBc particles than the conventional method. Our results demonstrated that the modified method produce a better yield and purity of HBc particles in an E. coli-expression system, which are fully characterised and suitable to be used for drug delivery applications.

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References
1.
Makrides S . Strategies for achieving high-level expression of genes in Escherichia coli. Microbiol Rev. 1996; 60(3):512-38. PMC: 239455. DOI: 10.1128/mr.60.3.512-538.1996. View

2.
Garcea R, Gissmann L . Virus-like particles as vaccines and vessels for the delivery of small molecules. Curr Opin Biotechnol. 2004; 15(6):513-7. DOI: 10.1016/j.copbio.2004.10.002. View

3.
Jana S, Deb J . Strategies for efficient production of heterologous proteins in Escherichia coli. Appl Microbiol Biotechnol. 2005; 67(3):289-98. DOI: 10.1007/s00253-004-1814-0. View

4.
Beterams G, Bottcher B, Nassal M . Packaging of up to 240 subunits of a 17 kDa nuclease into the interior of recombinant hepatitis B virus capsids. FEBS Lett. 2000; 481(2):169-76. DOI: 10.1016/s0014-5793(00)01927-x. View

5.
Sivashanmugam A, Murray V, Cui C, Zhang Y, Wang J, Li Q . Practical protocols for production of very high yields of recombinant proteins using Escherichia coli. Protein Sci. 2009; 18(5):936-48. PMC: 2771296. DOI: 10.1002/pro.102. View