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  • Title: Separation of heparin on Sepharose CL-4B in the presence of high concentrations of ammonium sulphate.
    Author: Uchiyama H, Fujimoto N, Sakurai K, Nagasawa K.
    Journal: J Chromatogr; 1984 Mar 30; 287(1):55-66. PubMed ID: 6725457.
    Abstract:
    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.
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