Human Prothrombin

Th. R Derleth; J. A Penner

doi:10.1055/s-0038-1654239

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1970; 24(03/04): 311-324
DOI: 10.1055/s-0038-1654239

Originalarbeiten — Original Articles — Travaux Originaux

Schattauer GmbH

Human Prothrombin

Chromatographic Separation from Normal and Deficient Plasmas^[*]

Th. R Derleth M. D.

¹University of Michigan Medical Center, Department of Internal Medicine, Simpson Memorial Institute, Ann Arbor, Michigan

,

J. A Penner M. S., M. D.

¹University of Michigan Medical Center, Department of Internal Medicine, Simpson Memorial Institute, Ann Arbor, Michigan

› Author Affiliations

Abstract

Summary

A method is presented for the chromatographic separation of human prothrombin from variable volumes of plasma. The procedure is simple, consisting of chromatography on ECTEOLA which provides a potent product containing factor IX and X activity sufficient for therapeutic use. Additional chromatography on DEAE or Sephadex G200 will produce a prothrombin of high specific activity which appears homogeneous by ultracentrifugation and electrophoresis on cellulose acetate, although multiple protein fractions can be identified on starch gel electrophoresis.

Human prothrombin prepared by this method migrates as an alpha ₂-globulin and does not appear to be altered electrophoretically or antigenically from its original state in the plasma. The characteristics of human prothrombin obtained from patients with deficiencies of factor VII or IX do not differ from normal.

Full Text

References

References
1 Seegers W. H, Loomis E. C, Vandenbelt J. M. Preparation of Prothrombin Product: isolation of prothrombin and its properties. Arch. Biochem 06: 85 1945;
2 Miller K. D. Chromatographic Isolation of Plasma Prothrombin and Trans-a-glucosylase. J. biol. Chem 231: 987 1958;
3 Lewis M. L, Ware A. G. Simple Procedure for Separation of Prothrombin and Accelerator Globulin from Citrated Human Plasma. Proc. Soc. exp. Biol. (N. Y) 84: 636 1953;
4 Alexander B. IVth International Congress of Biochemistry: Blood Clotting Factors. Ed. Deutsch E. Pergamon Press; New York: 1959
5 Lanchantin G. F, Friedmann J. A, De Groot J, Mehl J. W. Preparation of Human Plasma Prothrombin and Some of Its Sedimentation Properties. J. biol. Chem 238: 238 1963;
6 Lanchantin G. F, Friedmann J. A. Isolation of Human Plasma Prothrombin of High Specific Activity by Gel Filtration. Proc. Soc. exp. Biol. (N.Y) 114: 584 1963;
7 Shapiro S. S, Waugh D. F. The Purification of Human Prothrombin. Thrombos. Diathes. haemorrh. (Stuttg) 16: 469 1966;
8 Ware A. G, Seegers W. H. Two Stage Procedure for the Quantitative Determination of Prothrombin Concentration. Amer. J. clin. Path 19: 471 1949;
9 Quick A. J. Hemorrhagic Diseases. Lea and Fiebiger; Philadelphia, Pa: 1957
10 Penner J. A. Plasma Prothrombin Activation in Hemorrhagic Disorders. Amer. J. clin. Path 42: 561 1964;
11 Simson Jr. L, Oberman H, Penner J. A. Clinical Evaluation of Cryo Precipitated Factor VIII. Amer. med. Assoc 199: 554-558 1967;
12 Oberman H, Penner J. A. Utilization of the Residual Plasma Following Preparation of Factor VIII Cryoprecipitate. J.A.M. A 205: 819-821 1968;
13 Folin O, Giocalteu V. On Tyrosine and Tryptophane Determinations in Proteins. J. biol. Chem 73: 627 1927;
14 Svennerholm L. Sialic Acids and Derivative: Estimation by the Ion Exchange Method: Methods in Enzymology VI. Eds. Colowick S. P, Kaplan N. O. 459 Academic Press; New York: 1963
15 Morris D. L. Quantitative Determination of Carbohydrates with Dreywood’s Anthrone Reagent. Science 107: 254 1945;
16 Elson L. A, Morgan W. J. A Colorimetric Method for the Determination of Glucosamine and Chondrosamine. Biochem. J 27: 1824 1933;
17 Roseman S, Doffner I. Colorimetric Method for Determination of Glycosamine and Galactos- amine. Anal. Chem 28: 1743 1956;
18 Gelman Procedures, Techniques and Apparatus for Electrophoresis. Gelman Instrument Company. Ann Arbor; Michigan.:
19 Smithies O. Zone Electrophoresis in Starch Gels: Group Variations in the Serum Proteins of Normal Human Adults. Biochem. J 61: 629 1955;
20 Alfonso E. Quantitative Immunoelectrophoresis of Serum Proteins. Clin. chim. Acta 10: 114 1964;
21 Winzler R. J. Determinations of Serum Glycoproteins. Ed. Glick C. Interscience Publishers, Inc; N.Y: 1955. Vol.II, 279.
22 Miller K. D. Personal communication..
23 Richter K, Klingenberg K, Reinicke D. Blutplasmafraktionen zur Behandlung von Gerinnungsstörungen. Ges. inn. Med 23: 18 1968;
24 Tishkoff G. H, Williams L. C, Brown D. M. Preparation of Highly Purified Prothrombin Complex. J. biol. Chem 243: 4151 1968;
25 Lamy F, Waugh D. F. The Physical Changes of Prothrombin Under Various Experimental Conditions. Thrombos. Diathes. haemorrh. (Stuttg) 02: 188 1958;