MEMBRANE-BASED THIOL/DISULFIDE EXCHANGE

描述（改编自申请人的摘要）：蛋白质越来越多地 用作药物、诊断剂、生物化学中的催化剂 过程，和生长促进剂在大肠杆菌细胞培养。商业 涉及使用含巯基蛋白质的方法受到以下因素的阻碍 缺乏实际的方法来操纵的氧化态 蛋白质硫醇 虽然蛋白质硫醇的氧化态可以是 通过硫醇/二硫化物交换操作，常规方法需要 加入过量的昂贵的、潜在危险的硫醇试剂。 该计划旨在开发一种新型的膜基 硫醇/二硫化物交换方法克服了这些缺点。 的 该方法使用催化量的硫醇试剂， 使用基于膜的方法再生。 在第一阶段，研究人员 成功地证明了该工艺的可行性，使用它：1） 还原氧化型谷胱甘肽中的二硫键，和2）重新激活 一种被空气氧化失活的酶上的必需硫醇。 在第二阶段，研究人员将：1）描述过程，2） 证明该方法对三个目标应用的适用性， 3)确定扩大规模的问题，以及4）展示技术和经济 以中试规模生产目标产品的工艺的可行性 为第三阶段商业化做准备。

DESCRIPTION (Adapted from applicant's abstract):Proteins are increasingly being used as pharmaceuticals, diagnostic agents, catalysts in biochemical processes, and growth promoters in mammalian-cell culture. Commercial processes involving the use of thiol-containing proteins are hampered by the lack of practical processes to manipulate the oxidation state of protein thiols. Although the oxidation state of protein thiols can be manipulated by thiol/disulfide exchange, conventional processes require adding an excess amount of expensive, potentially hazardous thiol reagent. This program is aimed at the development of a novel membrane-based thiol/disulfide-exchange process that overcomes these drawbacks. The process uses a catalytic amount of thiol reagent that is continuously regenerated using a membrane-based process. In Phase I, the investigator successfully demonstrated feasibility of the process, using it: 1) to reduce the disulfide linkage in oxidized glutathione, and 2) to reactivate essential thiols on an enzyme that had been inactivated by air oxidation. In Phase II, the investigators will:1) characterize the process, 2) demonstrate the applicability of the process to three target applications, 3) identify scale-up issues, and 4) demonstrate the technical and economic feasibility of the process to produce a target product at pilot-plant scale in preparation for Phase III commercialization.