Purification of recombinant HBc antigen expressed in Escherichia coli and Pichia pastoris: comparison of size-exclusion chromatography and ultracentrifugation

Geminiviral vectors based on bean yellow dwarf virus for production of vaccine antigens and monoclonal antibodies in plants

“…19,24 Centrifugation methods, however, have inherent limitations in scalability and time efficiency, and thereby are not optimal for large-scale VLP production. 21,31 To improve the scalability and robustness of VLP purification procedures, the authors have developed a scalable purification process for Norwalk VLP based on low pH precipitation and conventional chromatographic steps. 21,25 In this procedure, cell debris and ribulose 1,5-bisphosphate carboxylaseoxygenase (RuBisCo), the most abundant plant host protein in leaves, are removed from the extract by precipitation and centrifugation under low pH buffer conditions (pH 5.3).…”

Effect of different operating modes and biomass concentrations on the recovery of recombinant hepatitis B core antigen from thermal-treated unclarified Escherichia coli feedstock

“…Various methods for the purification of HBcAg have been widely reported and these methods are dependent on the physical properties of HBcAg particles. Size exclusion (Rolland et al, 2001), packed bed chromatography (Tang et al, 2007), cesium chloride and sucrose gradient ultracentrifugation (Tan et al, 2003) are the common methods employed, but the latter remains the choice of recovery method and has been largely reported in literature. However, several solid-liquid separation and concentration steps are necessary for these conventional methods that contributed to a higher overall production cost, loss of desired product and a longer processing time.…”

Soluble expression and purification of hepatitis B core antigen (HBcAg) subgenotype B3 in Escherichia coli using thioredoxin fusion tag

“…in previous research in P. pastoris that required 4 days [9]. In this research we used E. coli BL21 (DE3) as the expression host.…”

“…Previous studies had successfully propagated and expressed HBcAg in prokaryote and eukaryote systems such as E. coli and Pichia pastoris, respectively [9]. The expression of HBcAg protein in E. coli has been done before; furthermore, this protein can be expressed in fusion form with other proteins such as the EDIII-2 protein, but it is still expressed in insoluble form.…”

“…Meanwhile, HBcAg expression in P. pastoris needs longer time with more complex cell lysis procedures [9]. Thus, the objective of this study was to construct HBcAg encoding gene HBV/B subgenotype B3 and to express the recombinant protein in E. coli system in soluble form.…”