Immunochemical Studies on Proteins Important in Blood Coagulation

Abstract

SummaryThis report draws attention to the fact that only minute quantities of purified serum proteins (1 —15 μg/kg body weight) are required to stimulate antibody production. Injection of amounts greater than 1 mg/kg either is a waste or favors the production of non-specific antisera. Potent antisera can be produced in rabbits with a single intramuscular injection using either Al(OH)3 gel or Amphojel as an adjuvant.A number of purified plasma protein preparations which appear homogenous by ultra centrifugation or biological activity studies actually contain several antigenic components and are immunologically heterogeneous, when checked with highly specific antisera.Fibrinogen prepared by alcohol fractionation and tannic acid purification was found to be free of prothrombin, antihemophilic factor and Profibrinolysin. Both Immunoelectrophoresis and double diffusion in agar indicated a homogeneous preparation.Fibrinogens prepared from human, canine and bovine plasmas exhibited a low species specificity.

Plasminogen-125I responses in dogs to a single injection of urokinase and typhoid vaccine and to vascular injury

“…The supernates were then saturated 30% with (NH4) 2SO4 at 3°C with a gentle mixing for 5 min and centrifuged at 3000 g at 3°C for 10 The size of the column was 2.5 X 70 cm and the fraction volume, 10 ml. The flow rate was about 150 ml/hr.…”

Congenital dysprothrombinemia: an inherited structural disorder of human prothrombin

“…The recent development of Received for publication 9 June 1969. methods for the purification of human prothrombin (1)(2)(3) and the availability of specific antisera (4)(5)(6)(7) have permitted the use of physical and immunologic techniques for the investigation of this problem. Thus, one previously reported case of congenital hypoprothrombinemia (8,9) has been found to lack almost completely any material capable of being fractionated by a standard technique for the isolation of plasma prothrombin (10).…”

Studies on the purification of antihemophilic factor (factor VIII)

“…Cryoprecipitated antihemophilic factor contained [14][15][16] stainable bands when subjected to electrophoresis on acrylamide gel (Fig. 3).…”