1993 Volume 22 Issue 2 Pages 75-84
Polyclonal and monoclonal antibodies to porcine elastase were produced and characterized. Production of monoclonal antibodies was performed by somatic cell fusion. Titers and cross reactivites of the antibodies were studied by enzyme-linked immunosorbent assay (ELISA). Each of these polyclonal (R90-03, R90-04) and monoclonal (MAb653F2) antibodies reacted in dose-dependent fashion with porcine elastase. The affinities, as estimated by cross-reactivities of polyclonal antibodies, were highest for porcine elastase and low for human elastase and trypsin. On the other hand, the affinity of the monoclonal antibody was highest for porcine elastase, intermediate for human elastase and lowest for trypsin. Using ELISA methods, with the antibody capture assay, the quantitative determination of porcine elastase was possible using polyclonal antibodies between the range from 0.8 to 80 ng/ml. The quantitative determination of human elastase by antibody capture assay was also possible with use of the monoclonal antibody between the range from 4 to 80 ng/ml. In addition, the presence of elastase in vascular wall smooth muscle cells was demonstrated and the secretion of smooth muscle cell-derived elastase in normal and arteriosclerotic conditions was studied with fluorescent antibody methods using one of the polyclonal antibodies (R90-04). Findings suggested that the secretion of elastase was markedly reduced in intimal smooth muscle cells in arteriosclerotic lesions. The antibodies produced and characterized in the present study may be useful for future pathophysiological investigation of elastase.
