RAPID FRACTIONATION OF BOVINE TRANSFERRIN USING IMMOBILIZED GANGLIOSIDES

Bovine transferrin (BTF) was fractionated from bovine whey using ganglioside affinity chromatography. After loading the immobilized matrix with a 2% whey solution, the matrix was washed with sodium acetate buffer at pH 4 containing1 M NaCl before elution of BTF with sodium phosphate buffers at pH 7....

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Bibliographic Details
Published in:Preparative biochemistry & biotechnology 2001-05, Vol.31 (2), p.89-102
Main Authors: Walsh, M. K., Nam, Seung-Hee
Format: Article
Language:English
Subjects:
Animals
Anion Exchange Resins - chemistry
Blotting, Western
Cattle
Chromatography, Affinity
Chromatography, Ion Exchange
Concanavalin A - chemistry
Electrophoresis, Polyacrylamide Gel
gangliosides
Gangliosides - chemistry
Immobilization
Milk - chemistry
Milk Proteins - chemistry
Milk Proteins - isolation & purification
Resins, Synthetic
Subcellular Fractions
transferrin
Transferrin - chemistry
Transferrin - isolation & purification
Whey Proteins
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Summary:Bovine transferrin (BTF) was fractionated from bovine whey using ganglioside affinity chromatography. After loading the immobilized matrix with a 2% whey solution, the matrix was washed with sodium acetate buffer at pH 4 containing1 M NaCl before elution of BTF with sodium phosphate buffers at pH 7. Concanavalin-A affinity and ion exchange chromatography were used for further purification. The ganglioside column showed a 74.2% BTF recovery from whey and BTF was enriched to 61% purity with ion exchange chromatography. Bovine transferrin was identified by SDS-PAGE and western analysis. The Concanavalin-A affinity and ion exchange chromatography steps enriched BTF in the samples and removed other whey proteins from ganglioside purified fractions. These results indicate that immobilized ganglioside can be used to fractionate BTF from bovine whey. Our novel ganglioside affinity chromatography is rapid and efficient for the fractionation of BTF from whey.
ISSN:1082-6068
1532-2297
DOI:10.1081/PB-100103375