Selective adsorption of immunoglobulins and glucosylated proteins on phenylboronate-agarose.
Aminophenylboronate-substituted agarose in 20 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulphonic acid, pH 8.5, selectively adsorbs immunoglobulins and complement factors C3 and C4 from human serum. The selectivity of binding is strongly influenced by the presence of magnesium chloride in the sample buffer. Adsorbed immunoglobulins are quantitatively eluted by sorbitol, but only partially by ethylene glycol or methylcellosolve. Aniline-agarose of a similar degree of substitution shows only weak adsorption of serum proteins under similar experimental conditions, thus indicating the important contribution of the boronate moiety to this interaction. Immunoglobulin adsorption seems not to be due to the cis-diol complexation used extensively for the chromatographic determination of non-enzymatically glucosylated proteins. Hydrophobic and pi-pi interactions with the aromatic structure of the ligand seem also to contribute to protein binding. The behaviour of aminophenylboronate-liganded agarose is, in some respects, rather similar to that of the so-called "thiophilic adsorbents".
Brena
B M
BM
Biochemical Separation Centre, Uppsala University, Uppsala Biomedical Centre, Sweden.
Batista-Viera
F
F
Rydén
L
L
Porath
J
J
D000327:Adsorption; D001798:Blood Proteins / Q000737:chemistry*; D001897:Boronic Acids / Q000737:chemistry*; D002853:Chromatography, Liquid; D003176:Complement C3 / Q000737:chemistry; D003181:Complement C4 / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005947:Glucose / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007136:Immunoglobulins / Q000737:chemistry*; D015636:Magnesium Chloride / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry*; D012709:Serum Albumin / Q000737:chemistry
D001798:Blood Proteins; D001897:Boronic Acids; D003176:Complement C3; D003181:Complement C4; D007136:Immunoglobulins; D012709:Serum Albumin; D015636:Magnesium Chloride; D012685:Sepharose; D005947:Glucose; C010686:benzeneboronic acid