Rapid isolation of human plasma anti-alpha-galactoside antibody using sugar-specific binding to guar galactomannan or agarose.
Indian J Biochem Biophys
;
1992 Jun; 29(3): 266-70
Article
in English
| IMSEAR
| ID: sea-26601
ABSTRACT
A method of purifying the naturally occurring antibody to alpha-galactoside moiety (anti-alpha-Gal) in human plasma by a single-step affinity chromatography on cross-linked guar galactomannan (CLGG) or agarose (Sepharose 4B) is described. IgG nature of the two preparations, as revealed by agar gel diffusion, as well as their preference for alpha-anomer of galactose, as revealed in inhibition of their agglutination of trypsinized rabbit erythrocytes by sugars, identified them with anti-alpha-Gal. The antibody binding capacity of Sepharose 4B was only a third of that of CLGG. Both gels showed similar dependence on ionic strength for binding. The pH optimum for binding of anti-alpha-Gal to CLGG was 8.0. Significantly anti-alpha-Gal binding to Sepharose was unaffected by CNBr activation and ligand coupling to the gel, thus warning that contaminating plasma could introduce artifacts in agarose-based chromatography of human tissue biomolecules.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Osmolar Concentration
/
Sepharose
/
Humans
/
Carbohydrates
/
Chromatography, Affinity
/
Galactosides
/
Hydrogen-Ion Concentration
/
Mannans
/
Antibodies
Language:
English
Journal:
Indian J Biochem Biophys
Year:
1992
Type:
Article
Similar
MEDLINE
...
LILACS
LIS