Immunochemical Studies of Plasma Kallikrein

Andranik Bagdasarian; Biswajit Lahiri; Richard C. Talamo; Pat Wong; Robert W. Colman

doi:10.1172/JCI107892

Immunochemical Studies of Plasma Kallikrein

Andranik Bagdasarian, Biswajit Lahiri, Richard C. Talamo, Pat Wong, and Robert W. Colman

Published December 1, 1974 - More info

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Abstract

A monospecific antibody against human plasma kallikrein has been prepared in rabbits with kallikrein further purified to remove gamma globulins. The antisera produced contained antikallikrein and also anti-IgG, in spite of only 8% contamination of kallikrein preparation with IgG. The latter antibody was removed by adsorption of antisera with either Fletcher factor-deficient plasma or with purified IgG. Both kallikrein and prekallikrein (in plasma) cross-react with the antibody with no apparent difference between the precipitation arcs developed during immunoelectrophoresis and no significant difference in reactivity when quantified by radial immunodiffusion.

Kallikrein antibody partially inhibits the esterolytic and fully inhibits the proteolytic activity of kallikrein. In addition, the antibody inhibits the activation of prekallikrein, as measured by esterase or kinin release. The magnitude of the inhibition is related to the molecular weight of the activator used. Thus, for the four activators tested, the greatest inhibition is observed with kaolin and factor XII_A, while large activator and the low molecular weight prekallikrein activators are less inhibited.

With the kallikrein antibody, the incubation of kallikrein with either plasma or partially purified C1 esterase inactivator results in a new precipitin arc, as detected by immunoelectrophoresis. This finding provides physical evidence for the interaction of the enzyme and inhibitor. No new arc could be demonstrated between kallikrein and α₂-macroglobulin, or α₁-antitrypsin, although the concentration of free kallikrein antigen decreases after interaction with the former inhibitor.

By radial immunodiffusion, plasma from healthy individuals contained 103±13 μg/ml prekallikrein antigen. Although in mild liver disease, functional and immunologic kallikrein are proportionally depressed, the levels of prekallikrein antigen in plasma samples from patients with severe liver disease remains 40% of normal, while the functional kallikrein activity was about 8%. These observations suggest that the livers of these patients have synthesized a proenzyme that cannot be converted to active kallikrein.

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