Parman A U, Rideout J M
J Chromatogr. 1983 Feb 4;256(2):283-91. doi: 10.1016/s0021-9673(01)88240-6.
A procedure has been developed for purification of porcine proinsulin by high-performance liquid chromatography from a preparation obtained as a side product during the Sephadex G-50 gel filtration of an impure porcine insulin preparation. Reversed-phase chromatography was carried out on octadecylsilica as the stationary phase with graded mixtures of acetonitrile or methanol-acetonitrile and phosphate buffer pH 2.4 as the mobile phase. The crude preparation separated into five different groups of proteins, the proinsulin-containing peak being identified by the co-eluting internal proinsulin marker. After purification by conventional procedures (separation, pooling, freeze drying, desalting, reprecipitation and drying) this peak fraction was rechromatographed by high-performance liquid chromatography (for final purification) to give a single peak protein which had identical electrophoretic mobility to that of commercial porcine proinsulin, and which converted to a protein with electrophoretic mobility similar to that of porcine insulin.
已开发出一种通过高效液相色谱法从在不纯猪胰岛素制剂的葡聚糖凝胶G-50凝胶过滤过程中作为副产物获得的制剂中纯化猪胰岛素原的方法。以十八烷基硅胶为固定相，以乙腈或甲醇-乙腈与pH 2.4的磷酸盐缓冲液的梯度混合物为流动相进行反相色谱。粗制剂分离成五组不同的蛋白质，含胰岛素原的峰通过共洗脱的内源性胰岛素原标记物来鉴定。通过常规程序（分离、合并、冷冻干燥、脱盐、再沉淀和干燥）纯化后，该峰馏分通过高效液相色谱进行再色谱（用于最终纯化），得到一种单一峰蛋白，其电泳迁移率与市售猪胰岛素原相同，并且转化为电泳迁移率与猪胰岛素相似的蛋白质。
