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.
authors.affiliation: Institute of Pharmaceutical Science, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Izzat Fahimuddin
authors.initials: IF
authors.lastname: Bin Mohamed Suffian
authors.affiliation: Randall Division of Cell &Molecular Biophysics, King's College London, New Hunt's House, London SE1 1UL, UK.
authors.forename: Mitla
authors.initials: M
authors.lastname: Garcia-Maya
authors.affiliation: Randall Division of Cell &Molecular Biophysics, King's College London, New Hunt's House, London SE1 1UL, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Brown
authors.affiliation: Biomolecular Spectroscopy Centre, King's College London, The Wolfson Wing, Hodgkin Building, SE1 1UL, UK.
authors.forename: Tam
authors.initials: T
authors.lastname: Bui
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Nishimura
authors.affiliation: Barts and The London School of Medicine and Dentistry, Queen Mary University of London, 4 Newark Street, London E1 2AT, UK.
authors.forename: Amir Rafiq Bin Mohammad Johari
authors.initials: AR
authors.lastname: Palermo
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Ogino
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
authors.affiliation: Institute of Pharmaceutical Science, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Khuloud T
authors.initials: KT
authors.lastname: Al-Jamal
mesh_terms: D015153:Blotting, Western; D002942:Circular Dichroism; D004337:Drug Carriers / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006512:Hepatitis B Core Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006515:Hepatitis B virus / Q000235:genetics* / Q000648:ultrastructure; D018625:Microscopy, Atomic Force; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure
chemical_list: D004337:Drug Carriers; D006512:Hepatitis B Core Antigens; D011994:Recombinant Proteins