Purification of human intrinsic factor using high‐performance ion‐exchange chromatography as the final step
- 6 May 1985
- journal article
- Published by Wiley in FEBS Letters
- Vol. 184 (1) , 14-19
- https://doi.org/10.1016/0014-5793(85)80643-8
Abstract
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.Keywords
This publication has 18 references indexed in Scilit:
- The preparation of human intrinsic factor-cobalamin complex from human gastric juice by immunoadsorptionClinica Chimica Acta; International Journal of Clinical Chemistry, 1984
- Effect of pancreatic extracts on the faecal excretion and on the serum concentration of cobalamin and cobalamin analogues in cystic fibrosisClinica Chimica Acta; International Journal of Clinical Chemistry, 1984
- Sequestration of crystalline and endogenous cobalamin by R binders down to the distal ileum in exocrine pancreatic dysfunctionClinica Chimica Acta; International Journal of Clinical Chemistry, 1983
- A simplified technique to isolate the porcine and human ileal intrinsic factor receptors and studies on their subunit structuresBiochemical and Biophysical Research Communications, 1978
- Primary structure of human intrinsic factor: progress report on cyanogen bromide fragmentationScandinavian Journal of Clinical and Laboratory Investigation, 1978
- Alteration of human intrinsic factor during affinity chromatography purification using concentrated guanidineFEBS Letters, 1977
- Clinical Chemistry: Characterization of R-Type Vitamin B12-Binding Proteins by Isoelectric FocusingScandinavian Journal of Clinical and Laboratory Investigation, 1975
- Analysis of monosaccharides by gas-liquid chromatography of the o-methyl glycosides as trifluoroacetate derivatives : Application to glycoproteins and glycolipidsJournal of Chromatography A, 1972
- Electrophoretic analysis of the major polypeptides of the human erythrocyte membraneBiochemistry, 1971
- Blockade of vitamin B12-binding sites in gastric juice serum and saliva by analogues and derivatives of vitamin B12 and by antibody to intrinsic factorBiochimica et Biophysica Acta (BBA) - General Subjects, 1967