Estimation Activity and Partial Purification of Ceruloplasmin from Sera of Patients with Chronic Renal Failure and Healthy Subjects

Abstract

CeruloplasminCP (EC1.16.3.1) activity has been assayed in (32) serum samples of patients with Chronic Renal Failure CRF, and (32) serum samples of healthy individuals without any clinically detectable diseases have been used as control group. The aim of this study is to measure CP activity and partially purifying the enzyme from sera of patients with CRF and healthy individuals. The results of this study revealed that CP activity of patient’s serum shows a highly significant decrease (p < 0.05) compared to that of the healthy individuals. Ceruloplasmin enzyme was partially purified from the serum of patient with CRF and and healthy individual by three steps which operates throughout under mild conditions 4°, acetate buffer (pH 6.8): First step including the use of ammonium sulfate precipitation, second step utilize dialysis with acetate buffer, while the third step consists of gel-filtration on Sephacryl S200(27×1.6cm) . Insufficiently purified ceruloplasmin fractions obtained at various stages are diluted to the appropriate ionic strength and re-utilized. Two proteinous components had been isolated by gel filtration chromatography from the supernatant produced by ammonium sulfate. According to Poly Acryl Amide Electrophoresis PAGE appear two bands by protein staining. Abbreviation: ceruloplasmin, chronic renal failure, ammonium sulfate, dialysis, electrophoresis, gel filtration.