Narayanan S R, Kakodkar S V, Crane L J
J. T. Baker Inc., Phillipsburg, New Jersey 08865.
Anal Biochem. 1990 Aug 1;188(2):278-84. doi: 10.1016/0003-2697(90)90606-a.
A high-performance affinity chromatography support based on silica has been developed for the immobilization of proteins containing primary amino groups. A hydrophilic polymer covalently bound to the silica surface minimizes nonspecific protein binding to the support while preserving high binding capacity. The Schiff base reaction involved in the coupling of a ligand to the affinity medium is rapid, allows the use of mild conditions during the coupling process, and results in a very stable linkage. Reaction parameters were studied for protein coupling to the affinity support to determine optimum binding conditions and dynamic capacity as a function of protein size. The stability of the ligand-matrix bond was determined. The performance and reproducibility of the affinity support are demonstrated by its use in the analysis of nitrophenyl sugar derivatives, purification of glycoproteins, and isolation of anti-bovine immunoglobulin G developed in rabbit.
一种基于硅胶的高性能亲和色谱载体已被开发用于固定含伯氨基的蛋白质。共价结合到硅胶表面的亲水性聚合物可最大限度减少蛋白质与载体的非特异性结合,同时保持高结合能力。配体与亲和介质偶联过程中涉及的席夫碱反应速度快,允许在偶联过程中使用温和条件,并产生非常稳定的连接。研究了蛋白质与亲和载体偶联的反应参数,以确定最佳结合条件以及作为蛋白质大小函数的动态容量。测定了配体 - 基质键的稳定性。该亲和载体在硝基苯基糖衍生物分析、糖蛋白纯化以及兔体内产生的抗牛免疫球蛋白G的分离中的应用证明了其性能和重现性。