Abstract
Hepatitis B core virus-like particles (HBc-VLP) have been widely used as carrier platforms to present an epitope of interest. Escherichia coli expression system is cost effective and produces high yields of recombinant protein. However major drawbacks include difficulties in obtaining soluble expression and tendency to form inclusion bodies. To boost solubility of proteins during expression of E. coli-derived HBc-VLPs carrying EBNA1 epitope, a statistical approach involving fractional factorial design and response surface methodology was used. For the first time, this approach was applied to quantitatively determine the impact of key parameters in shake-flask cultivation. Expression conditions including post-induction temperature and shaker-speed, and cell density at induction were optimized. Based on native agarose gel electrophoresis, optimized soluble protein cellular yield was 210.5 mg g−1 dry cell mass and volumetric yield was 272 mg L−1 of culture media. Findings highlight: 1) the significant interaction between post-induction temperature and shaker-speed on production, and; 2) sufficient oxygen level is required during induction. It is concluded that this statistical approach can be practically applied to optimize expression of HBc-VLP in shake-flask cultivation, and to determine key parameters for large-scale productions.
Published Version
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