Wu X, Linhardt R J
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242, USA.
Electrophoresis. 1998 Nov;19(15):2650-3. doi: 10.1002/elps.1150191514.
A new approach for separation, capillary affinity chromatography, is introduced for studying the interaction of heparin with antithrombin III and secretory leukocyte proteinase inhibitor. Heparin is covalently immobilized on the surface of an etched capillary through a silane spacer. The proteins are injected into the heparinized capillary, bound to the heparin, washed with buffer, eluted with sodium chloride in the same buffer using a pressure injection mode and eluting protein detected by absorbance. The resulting affinity separation is similar to that obtained from traditional affinity chromatography. The quantity of loaded protein in capillary affinity chromatography is at the nanogram level, offering an improvement over the milligram levels required for standard affinity chromatographic methods.
介绍了一种用于研究肝素与抗凝血酶III及分泌型白细胞蛋白酶抑制剂相互作用的新分离方法——毛细管亲和色谱法。肝素通过硅烷间隔物共价固定在蚀刻毛细管表面。将蛋白质注入肝素化毛细管中，使其与肝素结合，用缓冲液洗涤，然后在相同缓冲液中使用压力注射模式用氯化钠洗脱，通过吸光度检测洗脱的蛋白质。所得的亲和分离结果与传统亲和色谱法相似。毛细管亲和色谱法中加载蛋白质的量处于纳克水平，相较于标准亲和色谱法所需的毫克水平有了改进。
