Purification, characterization and development of radioimmunoassay of human liver ribonuclease

Abstract

Human liver ribonuclease (RNase) was purified 3600-fold into an electrophoretically homogeneous state by column chromatography on phosphocellulose, gel filtration, poly(G) affinity chromatography, and heparin affinity chromatography. The molecular weight of the RNase estimated by SDS disc electrophoresis was 19500. RNase was a heat- and pH-stable protein, and optimum activity was obtained at pH 7.0. The radioimmunoassay (RIA) for human liver RNase has been developed and the assay was shown to be sensitive (20 ng/ml), reproducible and specific. A good parallel relationship was observed between the standard curve and the dilution curves for serum and urine. No cross-reactivity was demonstrated between human liver and pancreatic RNase (less than 1%). In 44 normal subjects, the mean serum concentration of liver RNase determined by the RIA was found to be 99.4 ng/ml (SD ± 66.3).