Study of ceruloplasmin in renal insufficiency.
Ceruloplasmin, a copper carrier protein in mammal serum, was isolated by a variation of the Neo-Zgirski method consisting of a chromatography column in DEAE Sephadex A-50 gel, using different buffers for its selective elution. This protein was then precipitated with ammonium sulfate and an ethanol chloroform mixture. Finally, a dialysis against sodium acetate was performed and the samples were purified. In addition, the protein concentration and its characterization were determined. Enzyme kinetics were calculated using paraphenylenediamine as an artificial substrate, determined by the Michaelis constant. Furthermore, an SDS electrophoresis in polyacrylamide gel was performed, and the samples of porcine ceruloplasmin were compared with commercial bovine ceruloplasmin. Finally, a comparative study was carried out between the ceruloplasmin levels in serum from patients with chronic renal insufficiency (CRI) undergoing hemodialysis treatment in artificial kidneys with a cuprophan membrane, with a control group of donors. A similar study was performed with a group of patients suffering from the same illness, but undergoing dialysis treatment with polyacrylonitrile membrane. These results were expressed in mg% in relation to the ceruloplasmin isolated previously.
Martin Mateo
M C
MC
Department of Biochemistry Facultad de Ciencias, Universidad de Valladolid, Spain.
Maestro
J
J
D000085:Acetates; D019342:Acetic Acid; D000328:Adult; D000645:Ammonium Sulfate; D002570:Ceruloplasmin / Q000302:isolation & purification*; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007676:Kidney Failure, Chronic / Q000097:blood* / Q000628:therapy; D008297:Male; D008875:Middle Aged; D006435:Renal Dialysis
D000085:Acetates; D002570:Ceruloplasmin; D019342:Acetic Acid; D000645:Ammonium Sulfate