Bos O J, Fischer M J, Wilting J, Janssen L H
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, State University of Utrecht, The Netherlands.
J Chromatogr. 1988 Jan 22;424(1):13-21. doi: 10.1016/s0378-4347(00)81071-3.
For a thorough investigation of the drug-binding behaviour and other physicochemical properties of human serum albumin, one needs large amounts of specific fragments of albumin. Such fragments were obtained by careful proteolysis of the native protein with pepsin at pH 3.70. The fast protein liquid chromatographic technique was used to find the optimum experimental conditions for the separation of the fragments. By means of anion-exchange chromatography, chromatofocusing and gel permeation, it was possible to obtain a large fragment with a relative molecular mass of 46,000. The fragment could be assigned to segment 1-387 and therefore consists of domains 1 and 2 of the albumin structure. A 1-g amount of albumin produced 50 mg of a fragment that was 98% homogeneous.
为了全面研究人血清白蛋白的药物结合行为及其他物理化学性质，需要大量白蛋白的特定片段。通过在pH 3.70条件下用胃蛋白酶仔细消化天然蛋白质获得了这些片段。采用快速蛋白质液相色谱技术来寻找分离这些片段的最佳实验条件。借助阴离子交换色谱、色谱聚焦和凝胶渗透，有可能获得相对分子质量为46,000的大片段。该片段可归属于1 - 387区段，因此由白蛋白结构的结构域1和结构域2组成。1克白蛋白产生了50毫克纯度为98%的片段。
