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Purification of hog gastric intrinsic factor by a simple two-step procedure based on affinity chromatography and a selective guanidine hydrochloride gradient.
Gastroenterology. 1977 Jun; 72(6):1304-7.G

Abstract

Monocarboxylic acid derivaties of vitamin B12 were covalently coupled to 1,6-hexanediamine-substituted Sepharose by using a water-soluble carbodiimide resulting in 1.32 micronmoles of B12 coupled per ml of Sepharose. After a source of crude hog intrinsic factor (IF) was passed over the column, a selective linear gradient of guanidine HC1 (0 to 4.0 M) was used to remove IF and 4.0 to 7.5 M to elute NIF (a vitamin B12-binding glycoprotein not active in promoting vitamin B12 absorption). Anti-IF antibodies blocked 99% of the B12 binding by the isolated IF and only 1% of the B12 binding by NIF. Passage over a hydroxyapatite column resulted in IF 99% pure with a specific activity of 29.8 microng of B12 binding per mg of protein. IF so isolated exhibited one homogeneous band on polyacrylamide gel electrophoresis and corrected B12 malabsorption in a patient with pernicious anemia.

Authors

No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

870379

Citation

Francis, G L., et al. "Purification of Hog Gastric Intrinsic Factor By a Simple Two-step Procedure Based On Affinity Chromatography and a Selective Guanidine Hydrochloride Gradient." Gastroenterology, vol. 72, no. 6, 1977, pp. 1304-7.
Francis GL, Smith GW, Toskes PP, et al. Purification of hog gastric intrinsic factor by a simple two-step procedure based on affinity chromatography and a selective guanidine hydrochloride gradient. Gastroenterology. 1977;72(6):1304-7.
Francis, G. L., Smith, G. W., Toskes, P. P., & Sanders, E. G. (1977). Purification of hog gastric intrinsic factor by a simple two-step procedure based on affinity chromatography and a selective guanidine hydrochloride gradient. Gastroenterology, 72(6), 1304-7.
Francis GL, et al. Purification of Hog Gastric Intrinsic Factor By a Simple Two-step Procedure Based On Affinity Chromatography and a Selective Guanidine Hydrochloride Gradient. Gastroenterology. 1977;72(6):1304-7. PubMed PMID: 870379.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification of hog gastric intrinsic factor by a simple two-step procedure based on affinity chromatography and a selective guanidine hydrochloride gradient. AU - Francis,G L, AU - Smith,G W, AU - Toskes,P P, AU - Sanders,E G, PY - 1977/6/1/pubmed PY - 1977/6/1/medline PY - 1977/6/1/entrez SP - 1304 EP - 7 JF - Gastroenterology JO - Gastroenterology VL - 72 IS - 6 N2 - Monocarboxylic acid derivaties of vitamin B12 were covalently coupled to 1,6-hexanediamine-substituted Sepharose by using a water-soluble carbodiimide resulting in 1.32 micronmoles of B12 coupled per ml of Sepharose. After a source of crude hog intrinsic factor (IF) was passed over the column, a selective linear gradient of guanidine HC1 (0 to 4.0 M) was used to remove IF and 4.0 to 7.5 M to elute NIF (a vitamin B12-binding glycoprotein not active in promoting vitamin B12 absorption). Anti-IF antibodies blocked 99% of the B12 binding by the isolated IF and only 1% of the B12 binding by NIF. Passage over a hydroxyapatite column resulted in IF 99% pure with a specific activity of 29.8 microng of B12 binding per mg of protein. IF so isolated exhibited one homogeneous band on polyacrylamide gel electrophoresis and corrected B12 malabsorption in a patient with pernicious anemia. SN - 0016-5085 UR - https://www.unboundmedicine.com/medline/citation/870379/Purification_of_hog_gastric_intrinsic_factor_by_a_simple_two_step_procedure_based_on_affinity_chromatography_and_a_selective_guanidine_hydrochloride_gradient_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S001650857700184X DB - PRIME DP - Unbound Medicine ER -