Foot and mouth disease virus concentration and purification by affinity chromatography

Foot and mouth disease virus, (FMDV) from a crude cell lysate was purified in a single step by affinity chromatography with heparin as a ligand. The virus eluted from an Heparin-Ultrogel A4R column at 1M sodium chloride in 10 mM sodium phosphate buffer, pH 7.0, while most cell protein and albumin di...

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Bibliographic Details
Published in:Applied biochemistry and biotechnology 1996-12, Vol.61 (3), p.399-409
Main Authors: Canizo, A.A.N.D. (Facultad de Farmacia y Bioquimica, Buenos Aires, Argentina.), Mazza, M, Bellinzoni, R, Cascone, O
Format: Article
Language:English
Subjects:
Absorption spectra
Affinity
Affinity chromatography
Aphthovirus - isolation & purification
Biological and medical sciences
Buffers
CHROMATOGRAPHIE
Chromatography
Chromatography, Affinity
Chromatography, High Pressure Liquid
CROMATOGRAFIA
Foot & mouth disease
Foot-and-mouth disease virus
Fundamental and applied biological sciences. Psychology
Gel electrophoresis
Heparin
High performance liquid chromatography
Liquid chromatography
Microbiology
PURIFICACION
PURIFICATION
Purity
Sodium chloride
Sodium lauryl sulfate
Sodium phosphate
Spectrophotometry, Ultraviolet
Techniques used in virology
Ultracentrifugation
Virology
VIRUS FIEBRE AFTOSA
VIRUS FIEVRE APHTEUSE
Viruses
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Description
Summary:Foot and mouth disease virus, (FMDV) from a crude cell lysate was purified in a single step by affinity chromatography with heparin as a ligand. The virus eluted from an Heparin-Ultrogel A4R column at 1M sodium chloride in 10 mM sodium phosphate buffer, pH 7.0, while most cell protein and albumin did so at lower concentrations of sodium chloride in the same buffer. Purity of the eluted fraction containing the virus was assessed by SDS-PAGE, HPLC, ultracentrifugation, and UV absorption spectrum. With this method, intact viral particles are recovered in high yield (over 90%) and specific virus purity increases nearly 1000-fold. The capacity of the chromatographic matrix for the virus was found to be 1.1 mg viral mass per mL of hydrated gel
ISSN:0273-2289
1559-0291
DOI:10.1007/bf02787811