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.
Highlights
As the first VLP candidate and the first icosahedral VLP carriers, Hepatitis B Virus core (HBc) particles remain the most flexible and promising model for knowledge-based display of foreign peptide sequences[9,10]
wild type HBc (WT-HBc) particles were prepared by the “conventional” method (C) reported previously by others[26], Production steps involve bacterial culture, protein isolation, denaturation, purification and VLP assembly
We aimed at developing an improved method (I) for producing wild type HBc (WT-HBc) particles in high yields, suitable for drug delivery applications
Summary
As the first VLP candidate and the first icosahedral VLP carriers, HBc particles remain the most flexible and promising model for knowledge-based display of foreign peptide sequences[9,10]. HBc particles are hollow nanoparticles ranging from 30 to 34 nm in diameter with 7 nm thick envelopes[11] These core particles are icosahedral nucleocapsids with primarily triangulation number T = 3 or T = 4 symmetry, each containing 180–240 units of www.nature.com/scientificreports/. Escherichia coli offers a means for rapid, high yield and economical production of recombinant proteins[20,21]. High-level production of recombinant bio-nanoparticles in bacteria on a laboratory-scale is challenging. A yield optimisation study of HBc particles expression in E. coli is required to improve the productivity of the laboratory-scale for further drug delivery applications. This study focused on developing an improved method (I) for producing wild type HBc (WT-HBc) particles in high yields, suitable for drug delivery applications. We concluded that the modified method produced ~3 fold higher yield and greater purity of HBc particles than the conventional method
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