title: Heat treatment of unclarified Escherichia coli homogenate improved the recovery efficiency of recombinant hepatitis B core antigen.
abstract: Heat precipitation procedure has been regularly incorporated as a selective purification step in various thermostable proteins expressed in different hosts. This method is efficient in precipitation of most of the host proteins and also deactivates various host proteases that can be harmful to the desired gene products. In this study, introduction of heat treatment procedure in the purification of hepatitis B core antigen (HBcAg) produced in Escherichia coli has been investigated. Thermal treatment of the cell homogenate at 60 degrees C for 30 min prior to subsequent clarification steps has resulted in 1.4 times and 18% higher in purity and recovery yield, respectively, compared to the non-heat-treated cell homogenate. In direct capture of HBcAg by using anion-exchangers from unclarified feedstock, pre-conditioning the feedstock by heat treatment at 60 degrees C for 45 min has increased the recovery yield of HBcAg by 2.9-fold and 42% in purity compared to that treated for 10 min. Enzyme-linked immunosorbent assay (ELISA) analysis showed that the antigenicity of the core particles was not affected by the heat treatment process.
authors.affiliation: Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
authors.forename: Michelle Y T
authors.initials: MY
authors.lastname: Ng
authors.forename: Wen Siang
authors.initials: WS
authors.lastname: Tan
authors.forename: Norhafizah
authors.initials: N
authors.lastname: Abdullah
authors.forename: Tau Chuan
authors.initials: TC
authors.lastname: Ling
authors.forename: Beng Ti
authors.initials: BT
authors.lastname: Tey
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D011232:Chemical Precipitation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D006512:Hepatitis B Core Antigens / Q000302:isolation & purification*; D006358:Hot Temperature; D011994:Recombinant Proteins / Q000302:isolation & purification
chemical_list: D006512:Hepatitis B Core Antigens; D011994:Recombinant Proteins