title: Development of an enzyme-linked immunosorbent assay analytical platform for refolding yield determination of recombinant hepatitis B virus X (HBx) protein.
abstract: We report the development of a novel ELISA platform to quantitate hepatitis B virus X (HBx) protein refolding yields, which is critical for rational design and scaleup of aHBx bioprocess. HBx refolding yields were measured by determining the amount of HBx bound to immobilized GST-p53 on a "reduced glutathione"-functionalized maleimide surface. Refolding yields were distinguished from soluble yields, which were determined by measuring total HBx protein bound to a maleimide surface under reducing conditions. This platform is amenable to scaleup, and will expedite HBx production for structural and clinical studies, leading to the development of HBx-based therapy for liver cancer.
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore.
authors.forename: Anindya
authors.initials: A
authors.lastname: Basu
authors.forename: Susanna Su Jan
authors.initials: SS
authors.lastname: Leong
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008301:Maleimides / Q000737:chemistry / Q000378:metabolism; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000737:chemistry; D015534:Trans-Activators / Q000737:chemistry* / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000378:metabolism; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D008301:Maleimides; D011994:Recombinant Proteins; D015534:Trans-Activators; D016159:Tumor Suppressor Protein p53; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; C043592:maleimide