Dept. of Chemical Engineering, University of Colorado at Boulder, Boulder, CO 80309, USA.
Biotechnol Prog. 2010 May-Jun;26(3):743-9. doi: 10.1002/btpr.393.
We expressed recombinant murine growth hormone (rmGH) in E. coli as a cost-effective way to produce large quantities (gram scale) of the protein for use in murine studies of immunogenicity to therapeutic proteins. High hydrostatic pressure was used to achieve high solubility and high refolding yields of rmGH protein produced in E. coli inclusion bodies. A two-step column purification protocol was used to produce 99% pure monomeric rmGH. Secondary and tertiary structures of purified rmGH were investigated using circular dichroism and 2D-UV spectroscopy. The purified rmGH produced was found to be biologically active in hypophysectomized rats.
我们在大肠杆菌中表达重组鼠生长激素（rmGH），以经济有效的方式生产大量（克级）蛋白质，用于研究治疗性蛋白的免疫原性。高压处理用于提高大肠杆菌包涵体中表达的 rmGH 蛋白的高溶解度和高重折叠产率。两步柱纯化方案用于生产 99%纯的单体 rmGH。使用圆二色性和二维 UV 光谱研究了纯化的 rmGH 的二级和三级结构。发现纯化的 rmGH 在垂体切除大鼠中具有生物活性。
