Application of Continuous Flow Electrophoresis to the Study of the Blood Coagulation Proteins and the Fibrinolytic Enzyme System. I. Normal Human Materials1

Abstract

Continuous flow filter paper electrophoresis, as developed by Svensson and Brattsen (1), Grassmann and Hannig (2-4) and Durrum (5), offers a method of plasma fractionation in which protein denaturation appears minimal. This study involves the testing of such fractions for biological activity in various coagulation systems. With the exception of fibrinogen, the coagulation proteins are present in plasma in minute amounts which, at the present time, can be assayed only by their activities in specific clotting systems. Many of these coagulation factors are readily denatured; thus, rigid control with particular attention to temperature is necessary to obtain good recovery of biological activity.Previous electrophoretic studies concerning distribution of plasma coagulation proteins have employed paper electrophoresis with elutions from cut strips. Owen and McKenzie (6) found prothrombin to coincide with a2 globulin, proconvertin (stable conversion factor) with ft globulin, proaccelerin (labile conversion factor) in a variable area between y and f8 peaks, antithrombin between /3 and a2, and fibrinogen remaining at the point of application. Using the same technique, Frick and Hagen (7) found Hageman factor activity in a fraction located between ft and -y globulin. On the other hand, the electrophoretic mobility of purified prothrombin was described by Seegers, McClaughry, and Fahey (8) as approximately equivalent to that of a, globulin. The

A circulating anticoagulant in gamma-1A-multiple myeloma: its modification by penicillin.

“…The patient's plasma and a normal plasma were fractionated by starch block electrophoresis (23 (20,21). The initial 10 ml of plasma was separated into 28 fractions.…”

“…The test was then continued for an additional 6 minutes. E) To determine the inhibitory effect of isolated fractions, the patient's plasma was fractionated into 28 fractions by the continuous flow electrophoresis method of Lewis, Walters, Didisheim, and Merchant (20,21). For testing, a single normal donor served as the tontrol, and the same hemophiliac plasma was used throughout.…”

Studies on the Action of Hageman Factor: Evidence That Activated Hageman Factor in Turn Activates Plasma Thromboplastin Antecedent*

“…However, a profusion of experiments, beginning with those of Bordet and Gengou (33), demonstrated that glass and certain other substances can initiate or accelerate clotting in plasma depleted of its cells. One site of action of glass is Hageman factor, present in normal plasma lbut lacking in the plasma of patients with Hageman trait (2)(3)(4)(5)(6)(7)(8). Perhaps other factors are altered as well (7,34); it is difficult to distinguish between a direct effect of glass upon a clotting factor and an indirect effect mediated through its activation of Hageman factor.…”

Application of Continuous Flow Electrophoresis to the Study of the Blood Coagulation Proteins. Ii. The Hemophilias (A and B) *

“…The methods have been described previously (1,2). Plasmas were obtained from six patients, five of whom had suffered from severe and one (C.F.)…”

“…A previous communication (1) concerned the application of continuous flow electrophoresis to the fractionation of normal blood preparations and the study of the coagulation proteins in these fractions. It was considered of some interest to apply the same techniques to pathological plasmas obtained from patients suffering from congenital deficiencies of AHF (antihemophilic factor A) and PTC (plasma thromboplastin component, Christmas factor, antihemophilic factor B).…”