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Thromb Haemost 1979; 42(05): 1452-1459
DOI: 10.1055/s-0038-1657046
Original Article
Schattauer GmbH Stuttgart

Separation of Heparin into Subtractions by DEAE-Cellulose Chromatography

Robert H Yue
The Cardiovascular Research Division, Institute of Rehabilitation Medicine, New York University Medical Center, New York, U.S.A.
,
Toby Starr
The Cardiovascular Research Division, Institute of Rehabilitation Medicine, New York University Medical Center, New York, U.S.A.
,
Menard M Gertler
The Cardiovascular Research Division, Institute of Rehabilitation Medicine, New York University Medical Center, New York, U.S.A.
› Author Affiliations
Further Information

Publication History

Received 06 March 1979

Accepted 21 May 1979

Publication Date:
23 August 2018 (online)

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Summary

Commercial porcine heparin can be separated into three distinct subtractions by using DEAE-cellulose chromatography and a stepped salt gradient. Gram quantities of heparin can be fractionated by this technique. All three heparin subtractions can accelerate the inhibition of thrombin by antithrombin III with different efficiency. The specific activities of the high activity heparin, intermediate activity heparin and low activity heparin are 228 units/mg, 142 units/mg and 95 units/mg, respectively. Both the uronic acid content and the quantity of N-SO4 for all three heparin subfractions have been evaluated. The high activity heparin has the lowest uronic acid and N-SO4 content. The successful separation of commercial heparin into three distinct subfractions by means of ion-exchange chromatography suggests that the net charge on these three heparin components will serve as a model system in the elucidation of the structure and activity relationship to the biological function of heparin.

 

  • References

  • 1 Andersson LO, Barrowcliffe TW, Holmer E, Johnson EA, Sims GE C. 1976; Anticoagulant Properties of Heparin Fractionated by Affinity Chromatography on Matrix-Bound Antithrombin III and by Gel Filtration. Thrombosis Research 9: 575
    CrossrefPubMedGoogle Scholar
  • 2 Bitter T, Muir HM. 1962; A Modified Uronic Acid Carbazole Reaction. Analytical Biochemistry 4: 330
    CrossrefPubMedGoogle Scholar
  • 3 Cifonelli JA, King J. 1972; The Distribution of 2-Acetamido-2-deoxy-D-glucose Residues in Mammalian Heparins. Carbohydrate Research 21: 173
    CrossrefPubMedGoogle Scholar
  • 4 Cleland RL, Cleland MC, Lipsky JJ, Lyn VE. 1968; Ionic Polysaccharides. I. Adsorption and Fractionation of Polyelectrolytes on (Diethylamino) ethyl Cellulose. Journal of American Chemical Society 90: 3141
    CrossrefPubMedGoogle Scholar
  • 5 Hook M, Bjork I, Hopwood J, Lindahl U. 1976; Anticoagulant Activity of Heparin: Separation of High-Activity and Low-Activity Heparin Species by Affinity Chromatography on Immobilized Antithrombin. Federation of Experimental Biology Society Letters 66: 90
    PubMedGoogle Scholar
  • 6 Jansson L, Ogren S, Lindahl U. 1975; Macromolecular Properties and End-Group Analysis of Heparin Isolated from Bovine Liver Capsule. Biochemical Journal 145: 53
    CrossrefPubMedGoogle Scholar
  • 7 Jaques LB, Bell HJ. 1959; Determination of Heparin. Methods of Biochemical Analysis 7: 253
    PubMedGoogle Scholar
  • 8 Lam LH, Silbert JE, Rosenberg RD. 1976; The Separation of Active and Inactive Forms of Heparin. Biochemical and Biophysical Research Communication 69: 570
    CrossrefPubMedGoogle Scholar
  • 9 Lane DA, MacGregor IR, Michalski R, Kakkar VV. 1978; Anticoagulant Activities of Four Unfractionated and Fractionated Heparins. Thrombosis Research 12: 257
    CrossrefPubMedGoogle Scholar
  • 10 Lasker SE, Shvala SS. 1966; Physiochemical Studies of Fractionated Bovine Heparin. I. Some Dilute Solution Properties. Archives of Biochemistry and Biophysics 115: 360
    CrossrefPubMedGoogle Scholar
  • 11 Lindahl U, Cifonelli JA, Lindahl B, Roden L. 1965; a The Role of Serine in the Linkage of Heparin to Protein. Journal of Biological Chemistry 240: 2821
    PubMedGoogle Scholar
  • 12 Noltmann EA, Gubler CG, Kuby SA. 1961; Glucose 6-Phosphate Dehydrogenase (Zwischenferment) I. Isolation of the Crystalline Enzyme from Yeast. Journal of Biological Chemistry 236: 1225
    PubMedGoogle Scholar
  • 13 Ratliff RL, Weaver RH, Lardy HA, Kuby SA. 1964; Nucleoside Triphosphate- Nucleoside Diphosphate Transphosphorylase (Nucleoside Diphosphokinase). I. Isolation of the Crystalline Enzyme from Brewer’s Yeast. Journal of Biological Chemistry 239: 301
    PubMedGoogle Scholar
  • 14 Ringertz NR, Reichard P. 1960; Chromatography of Sulfate Containing Mucopolysaccharides. Acta Chemica Scandinavica 14: 303
    CrossrefPubMedGoogle Scholar
  • 15 Robinson HC, Horner AA, Hook M, Ogren S, Lindahl U. 1978; A Proteoglycan Form of Heparin and its Degradation to Single-chain Molecules. Journal of Biological Chemistry 253: 6687
    PubMedGoogle Scholar
  • 16 Yue RH, Starr T, Gertler MM. 1973; Quantitative Determination of Total Antithrombin III in Plasma. Thrombosis et Diathesis Haemorrhagica 30: 84
    PubMedGoogle Scholar
  • 17 Yue RH, Starr T, Gertler MM. 1976; Isolation and Partial Characterization of Bovine Antithrombin III. Federation Proceedings 35: 322
    PubMedGoogle Scholar
  • 18 Yue RH, Starr T, Gertler MM. 1978; Chromatographic Behavior of Heparin. Circulation 58: 11-136
    PubMedGoogle Scholar
  • 19 Yue RH, Starr T, Gertler MM. 1979; Preparation of High Activity Heparin by DEAE- Cellulose Chromatography. Federation Proceedings 38: 1064
    PubMedGoogle Scholar