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 fractionated
into seven precipitates by cold ethanol precipitation. The yield (amount) of proteins in the
precipitates 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 total
amount of protein obtained was 3180.00 mg. The precipitates were fractionated into 25
fractions each by Anion Exchange Chromatography (AEC) and the amount of protein in
each fraction was obtained by Bradford protein assay. SDS-PAGE analysis was performed
on the fractions with proteins and their estimated molecular weights were obtained with the
aid of the molecular weight marker. The result indicated that cold ethanol precipitation and
anion exchange chromatographic techniques are valuable tools for the purification of
bovine 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; Globulin

DOI: 10.3126/jncs.v26i0.3624

Journal of Nepal Chemical Society
Vol. 26, 2010
Page: 2-12