Antibodies as Specific Renin Inhibitors: Studies with Polyclonal and Monoclonal Antibodies and Fab Fragments

Abstract

The use of antirenin antibody and Fab may provide a more specific physiologic tool and potential therapeutic agent than the existing pharmacologic inhibitors. The antibody combining site, by virtue of its larger size than organic compounds, has the capacity for a larger number of intramolecular contacts with its ligands, thus allowing increased selectivity and affinity. Renin-specific Fab obtained through immunization with purified dog renal renin has been studied. Fab had no effect on blood pressure in the sodium replete conscious dog but induced systemic hypotensive responses in the sodium deplete animal or during acute renovascular hypertension. These responses were accompanied with complete suppression of plasma renin and angiotensin II levels. The vasodepressor responses of Fab were comparable to the converting enzyme inhibitor, teprotide. However, the latter induced a greater renal vasodilator effect. Since antibodies and their fragments derived from immune sera are limited in their application to physiologic study with respect to lack of homogeneity, reproducibility and limitation of quantity, we obtained monoclonal antibodies to purified human renal renin. Fusion HR3 yielded 14 renin-antibody secreting clones. Six clones have been isolated and characterized. Isotypes include IgG1 and IgM. Kd's range from 6 × 10−9 to 7.5 × 10−10M renin concentration. Five clones produce antibodies with antienzymatic activity. IC50's of these antibodies range from 1 × 10−6 to 6 × 10−8M. All antibodies are renin-specific and do not bind several nonrenin enzymes and proteins examined. They are also highly species specific, i.e., do not crossreact with mouse, dog, bovine or rat renins but recognize hog renin. These catalytic-site directed antibodies may be highly potent and specific tools for physiologic studies in subhuman primates and man.