Separation of heparin on Sepharose CL-4B in the presence of high concentrations of ammonium sulphate.
Heparin was fractionated by chromatography on Sepharose CL-4B/3.8-2.0 M ammonium sulphate in 0.01 M hydrochloric acid at 4 degrees C based on a principle different from that of gel filtration--possibly due to multiple interaction mechanisms including those based on hydrophobic bonds. The results of the fractionation were very similar to those of chromatography on Phenyl-Sepharose CL-4B/3.8-2.0 M ammonium sulphate in 0.01 M hydrochloric acid at room temperature. That is, the separation was related to the molecular size distribution, N-acetyl content and anticoagulant activities. As the result of studies on a set of Sepharose 4B gels with different degrees of cross-linking, it has been shown that the introduction of the cross-linked structure, -O-CH2-CH(OH)-CH2-O-, into the gel matrix enhances the interaction between the Sepharose CL-4B gel and heparin, indicating that heparin retention by the highly cross-linked Sepharose 4B gel surpasses that by Phenyl-Sepharose CL-4B.
D000645:Ammonium Sulfate; D000818:Animals; D000925:Anticoagulants; D000990:Antithrombin III; D002845:Chromatography / Q000379:methods; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D003432:Cross-Linking Reagents; D005340:Fibrinogen / Q000302:isolation & purification; D006493:Heparin / Q000302:isolation & purification* / Q000494:pharmacology; D012685:Sepharose / Q000031:analogs & derivatives; D013552:Swine
D000925:Anticoagulants; D003432:Cross-Linking Reagents; C035771:Sepharose CL 4B; D000990:Antithrombin III; D005340:Fibrinogen; D006493:Heparin; D012685:Sepharose; D000645:Ammonium Sulfate