Thiophilic adsorption revisited.
Specific and efficient selection of serum immunoglobulins, but not other proteins, on T-gel remains difficult. T-gel capacity was determined for different activation conditions and serum loadings. Mass spectrometry analysis was used to identify the proteins found in the flow-through and in the eluted fractions. Alpha-2-macroglobulin and albumin were the major contaminants of the eluates. The influence of the competition between immunoglobulins and the other serum proteins on the adsorption was also studied. Using a serum depleted in immunoglobulins (flow-through of a first chromatography on T-gel), many serum proteins were retained on the T-gel, including albumin. We conclude that T-gel selectivity is less than absolute and may reflect for a large part the experimental conditions of the adsorption.
Hardouin
Julie
J
Laboratory of Protein Biochemistry and Proteomics, UMR CNRS 7033 (BioMoCeTi), UFR SMBH, Paris13 University, 93017 Bobigny cedex, France.
Duchateau
Magalie
M
Canelle
Ludovic
L
Vlieghe
Céline
C
Joubert-Caron
Raymonde
R
Caron
Michel
M
D000327:Adsorption; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D007136:Immunoglobulins / Q000302:isolation & purification*; D007163:Immunosorbent Techniques; D013438:Sulfhydryl Compounds
D007074:Immunoglobulin G; D007136:Immunoglobulins; D013438:Sulfhydryl Compounds