Purification of human complement factor D from the peritoneal fluid of patients on chronic ambulatory peritoneal dialysis.
We describe a simple procedure for the purification of human factor D from the peritoneal fluid (PF) of patients with end stage renal failure (ESRF) on chronic ambulatory peritoneal dialysis (CAPD). The main advantages of this method are: (1) a relative enrichment of factor D in PF versus plasma (factor D/total protein enriched 3.8-fold); thus, added to the elevated concentration of factor D in ESRF, the enrichment compared to normal human serum is approximately 50 fold. (2) This biological source of factor D is almost unlimited, since around 10 liters of PF are removed per day from each patient. The purification is performed in three simple steps-Bio Rex 70, Heparin Sepharose CL 6B and Mono S FPLC- and milligrams of highly purified factor D are obtained. Peritoneal fluid might be a valuable source for the purification of other low MW proteins which accumulate in renal failure.
Catana
E
E
Department of Medicine, Hôpital Cantonal Universitaire, Geneva, Switzerland.
Schifferli
J A
JA
D000911:Antibodies, Monoclonal; D001202:Ascitic Fluid / Q000737:chemistry*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D011416:Complement Factor D / Q000276:immunology / Q000302:isolation & purification*; D015941:Complement Hemolytic Activity Assay; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007676:Kidney Failure, Chronic / Q000378:metabolism / Q000628:therapy; D010531:Peritoneal Dialysis, Continuous Ambulatory
D000911:Antibodies, Monoclonal; C493488:CFD protein, human; D011416:Complement Factor D