Comprehensive study on binding capacity of human immunoglobulin G to Avid AL affinity gel.
Avid AL is an affinity chromatographic gel based on a synthetic ligand which has been designed for the purification of immunoglobulins. The binding capacities of the gel for human IgG were investigated under different conditions; various salts; salt concentrations; different buffer systems with pH ranging from 3.8 to 10.0; and, tissue culture supernatant supplemented with 20% serum. At a similar ionic strength, the binding capacity changed according to the salting out effect of the ions used. Kinetic parameters of the affinity adsorption showed that a strong salting out effect significantly decreased the dissociation constant. Avid AL gel purified IgG from human serum with a capacity of 26.2 mg/ml gel using binding buffer containing sodium sulfate. Recombinant Protein A gel was used as a control. In this study it also exhibited enhanced binding capacity with a high salt concentration binding buffer.
Shi
J Y
JY
Unisyn Technologies, Inc., Tustin, CA 92680.
Goffe
R A
RA
D001386:Azides; D002846:Chromatography, Affinity; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000302:isolation & purification / Q000378:metabolism*; D010637:Phenolsulfonphthalein; D011485:Protein Binding; D011725:Pyridines; D012492:Salts; D012685:Sepharose / Q000031:analogs & derivatives*; D019810:Sodium Azide
C076342:Avid AL gel; D001386:Azides; D007074:Immunoglobulin G; D011725:Pyridines; D012492:Salts; D012685:Sepharose; D019810:Sodium Azide; D010637:Phenolsulfonphthalein