Isolation of Plasma Proteins from Bovine Blood by Cold Ethanol Precipitation and Anion Exchange Chromatography

Abstract

Plasma (100.00 ml) obtained from bovine blood by centrifugation was fractionatedinto seven precipitates by cold ethanol precipitation. The yield (amount) of proteins in theprecipitates calculated from the standard curve of BSA was 1160.83 mg, 806.57 mg,1149.94 mg, 8.79 mg, 19.88 mg, 21.98 mg, and 13.97 mg respectively, while the totalamount of protein obtained was 3180.00 mg. The precipitates were fractionated into 25fractions each by Anion Exchange Chromatography (AEC) and the amount of protein ineach fraction was obtained by Bradford protein assay. SDS-PAGE analysis was performedon the fractions with proteins and their estimated molecular weights were obtained with theaid of the molecular weight marker. The result indicated that cold ethanol precipitation andanion exchange chromatographic techniques are valuable tools for the purification ofbovine blood to obtain high grade α, β and γ-fibrinogen, IgM (μ-globulin), IgG (γ -globulin),alpha (α –globulin), β-globulin (E-globulin) and albumin.Keywords: Plasma; Chromatography; Electrophoresis; Albumin; Fibrinogen; GlobulinDOI: 10.3126/jncs.v26i0.3624Journal of Nepal Chemical SocietyVol. 26, 2010Page: 2-12