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

Izzat Fahimuddin Bin Mohamed Suffian; Mitla Garcia-Maya; Paul Brown; Tam Bui; Yuya Nishimura; Amir Rafiq Bin Mohammad Johari Palermo; Chiaki Ogino; Akihiko Kondo; Khuloud T. Al-Jamal

doi:10.1038/srep43160

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

Izzat Fahimuddin Bin Mohamed Suffian, Mitla Garcia-Maya, Paul Brown, Tam Bui, Yuya Nishimura, Amir Rafiq Bin Mohammad Johari Palermo, Chiaki Ogino, Akihiko Kondo, Khuloud T. Al-Jamal^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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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.

Original language	English
Article number	43160
Journal	Scientific Reports
Volume	7
DOIs	https://doi.org/10.1038/srep43160
Publication status	Published - 3 Mar 2017

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title = "Yield Optimisation of Hepatitis B Virus Core Particles in E. coli Expression System for Drug Delivery Applications",

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.",

author = "{Bin Mohamed Suffian}, {Izzat Fahimuddin} and Mitla Garcia-Maya and Paul Brown and Tam Bui and Yuya Nishimura and Palermo, {Amir Rafiq Bin Mohammad Johari} and Chiaki Ogino and Akihiko Kondo and Al-Jamal, {Khuloud T.}",

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AU - Bin Mohamed Suffian, Izzat Fahimuddin

AU - Garcia-Maya, Mitla

AU - Brown, Paul

AU - Bui, Tam

AU - Nishimura, Yuya

AU - Palermo, Amir Rafiq Bin Mohammad Johari

AU - Ogino, Chiaki

AU - Kondo, Akihiko

AU - Al-Jamal, Khuloud T.

PY - 2017/3/3

Y1 - 2017/3/3

N2 - 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.

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

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