Reyl-Desmars F, Le Roux S, Linard C, Benkouka F, Lewin M J
Unité de Recherche de Gastroentérologie, I.N.S.E.R.M. U10, Hôpital Bichat, Paris.
C R Acad Sci III. 1989;308(9):251-4.
This communication reports the isolation and the purification of the gastric somatostatin receptor from the human cell line HGT-1. The receptor has been extracted from the cell membrane by Triton X 100, and a monoclonal antibody to this was prepared. A series of affinity chromatographies (Sepharose-antibody and Sepharose-somatostatin-14) and a final purification by steric exclusion on high performance liquid chromatography columns (HPLC) allowed us to obtain a fraction enriched 20,000 fold in 125I-Tyrll-somatostatin-14 specific binding (apparent dissociation constant: 7.6 x 10(-8) M). This fraction corresponded to a molecular mass of about 90 kDa (in presence of detergent) and to a maximal binding capacity of more than 10,000 pmol/protein. It therefore has a theoretical homogeneity close to 100%.
本通讯报道了从人细胞系HGT-1中分离和纯化胃生长抑素受体的过程。该受体已用Triton X 100从细胞膜中提取出来，并制备了针对此受体的单克隆抗体。一系列亲和色谱法（琼脂糖-抗体和琼脂糖-生长抑素-14）以及最后通过高效液相色谱柱（HPLC）进行空间排阻纯化，使我们获得了一个在125I-Tyr11-生长抑素-14特异性结合方面富集了20000倍的组分（表观解离常数：7.6×10⁻⁸ M）。该组分的分子量约为90 kDa（在有去污剂存在的情况下），最大结合容量超过10000 pmol/蛋白。因此，其理论纯度接近100%。
