Purification of human intrinsic factor using high‐performance ion‐exchange chromatography as the final step

FEBS Letters

Nicolas, Jean-Pierre¹  Masson, Catherine¹  Gueant, Jean-Louis¹  Gräsbeck, Ralph³  Michalski, Jean-Claude²  Kouvonen, Ilkka³ 

[1] Laboratoire de Biochimie Médicale, Faculté de Médecine, Université de Nancy I, BP 184, 54505 Vandæuvre-les-Nancy Cedex, France;Laboratoire de Chimie Biologique (Laboratoire associé au CNRS n°. 217), 59655 Villeneuve d'Ascq Cedex, France;Minerva Institute for Medical Research, POB 819, 00101 Helsinki 10, Finland

关键词: Intrinsic factor;    Cobalamin;    Vitamin B12 binding protein;    High-performance liquid chromatography Ion-exchange chromatography;    Glycoprotein;   

DOI  :  10.1016/0014-5793(85)80643-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Human intrinsic factor was purified 1430-fold from gastric juice with a yield of 75% using two steps: labile ligand affinity chromatography and high-performance ion-exchange chromatography. Intrinsic factor precipitated in the presence of specific autoantibodies and 15% sodium sulfate, had an estimated M _r of 59000 in 5% SDS electrophoresis and could bind to the specific ileal receptor in vitro. Its carbohydrate composition could be related to N-lactosaminic and O-glycosidic chains. High-performance ion-exchange chromatography was a mild, rapid and efficient procedure to separate completely intrinsic factor from haptocorrin (another glycoprotein of gastric juice which binds cobalamin) and from other contaminating proteins.

【 授权许可】

Unknown