School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore.
Anal Biochem. 2011 Nov 1;418(1):155-7. doi: 10.1016/j.ab.2011.07.014. Epub 2011 Jul 22.
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.
我们开发了一种新型 ELISA 平台来定量检测乙型肝炎病毒 X（HBx）蛋白的重折叠产率，这对于合理设计和放大 HBx 生物工艺至关重要。HBx 重折叠产率通过测定与固定在“还原型谷胱甘肽”功能化马来酰亚胺表面上的 GST-p53 结合的 HBx 量来确定。重折叠产率与可溶产率区分开来，后者通过在还原条件下测定结合在马来酰亚胺表面上的总 HBx 蛋白来测量。该平台适合放大，将加快 HBx 的生产用于结构和临床研究，从而开发基于 HBx 的肝癌治疗方法。
