Frantzen F, Grimsrud K, Heggli D E, Sundrehagen E
Iris Biotech AS, Mørkved, Norway.
J Chromatogr B Biomed Appl. 1995 Aug 4;670(1):37-45. doi: 10.1016/0378-4347(95)00141-5.
Different methods for covalent linkage of phenylboronic acid (PBA) to structural proteins and enzymes are presented. Protein-PBA conjugates, free in solution or immobilised on magnetizable polymer particles, were tested for their binding of D-sorbitol, D-mannose and glycohemoglobin (GHb). Similarly, alkaline phosphatase-PBA conjugates were used in an attempted enzyme-linked sorbent assay for the detection of GHb. Affinity chromatography on immobilised D-mannose and gel chromatographic studies of protein-PBA complexes with [14C]sorbitol, clearly illustrated a low affinity of the interaction studied. Glycated hemoglobin could not be detected using the enzyme-linked sorbent assay approach. However, GHb was found to be specifically retained on columns filled with protein-PBA-coated particles as affinity matrix, enabling the glycation level of blood samples to be determined.
本文介绍了将苯硼酸（PBA）共价连接到结构蛋白和酶上的不同方法。对溶液中游离或固定在可磁化聚合物颗粒上的蛋白质-PBA缀合物进行了测试，以检测它们与D-山梨醇、D-甘露糖和糖化血红蛋白（GHb）的结合情况。同样，碱性磷酸酶-PBA缀合物被用于尝试通过酶联吸附测定法检测GHb。固定化D-甘露糖上的亲和色谱以及蛋白质-PBA复合物与[14C]山梨醇的凝胶色谱研究清楚地表明，所研究的相互作用亲和力较低。使用酶联吸附测定法无法检测到糖化血红蛋白。然而，发现GHb可特异性保留在填充有蛋白质-PBA包被颗粒作为亲和基质的柱上，从而能够测定血样的糖化水平。
