title: Negative chromatography purification of hepatitis B virus-like particles using poly(oligo(ethylene glycol) methacrylate) grafted cationic adsorbent.
abstract: Poly(oligo(ethylene glycol) methacrylate) (POEGMA), an inert polymer was grafted onto an anion exchange adsorbent for the exclusion of relatively larger hepatitis B virus-like particles (HB-VLPs) from the anion exchange ligand (Q) and at the same time this process allowed the selective adsorption of smaller size Escherichia coli host cell proteins (HCPs). The chain lengths of the POEGMA grafted were modulated by varying the amount of monomers used in the polymer grafting. The purification factor and yield of the HB-VLPs obtained from the flow-through of negative chromatography were 2.3 and 66.0±3.1%, respectively, when shorter chain length of POEGMA (SQ) was grafted. Adsorbent grafted with longer chain of POEGMA (LQ) excluded some HCPs that are larger in size together with the HB-VLPs, reducing the purity of the recovered HB-VLPs. Further heat-treatment of the flow-through pool from SQ followed by centrifugation increased the purity of heat stable HB-VLPs to 87.5±1.1%. Heat-treatment of the flow through sample resulted in thermal denaturation and aggregation of HCPs, while the heat stable HB-VLPs still remained intact as observed under a transmission electron microscope. The performance of the negative chromatography together with heat treatment in the purification of HB-VLPs is far better than the reported bind-and-elute techniques.
authors.affiliation: Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia.
authors.forename: Micky Fu Xiang
authors.initials: MF
authors.lastname: Lee
authors.affiliation: Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia; Advanced Engineering Platform, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia.
authors.forename: Eng Seng
authors.initials: ES
authors.lastname: Chan
authors.affiliation: Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia; Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
authors.forename: Wen Siang
authors.initials: WS
authors.lastname: Tan
authors.affiliation: Department of Chemical Engineering, Waterloo Institute for Nanotechnology, University of Waterloo, 200 University Avenue West, Waterloo, ON, Canada N2L 3G1.
authors.forename: Kam Chiu
authors.initials: KC
authors.lastname: Tam
authors.affiliation: Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia; Advanced Engineering Platform, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia. Electronic address: tey.beng.ti@monash.edu.
authors.forename: Beng Ti
authors.initials: BT
authors.lastname: Tey
mesh_terms: D000327:Adsorption; D002852:Chromatography, Ion Exchange / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006512:Hepatitis B Core Antigens / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006515:Hepatitis B virus / Q000378:metabolism*; D008024:Ligands; D008689:Methacrylates / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D011109:Polymethacrylic Acids
chemical_list: D006512:Hepatitis B Core Antigens; D008024:Ligands; D008689:Methacrylates; D011109:Polymethacrylic Acids; C528061:poly(poly(ethylene glycol)methacrylate); D011092:Polyethylene Glycols
keywords: Heat-treatment; Hepatitis B virus core antigen (HBcAg); Negative chromatography; Poly(oligo(ethylene glycol) methacrylate) (POEGMA); Virus-like particles (VLPs)