High-performance affinity chromatography of human serum concanavalin A binding proteins.
A high-performance concanavalin A (Con A) affinity column Gelpack GL-L55C (Hitachi Kasei Industries) was successfully used for the fractionation of human serum Con A-binding proteins. Serum proteins that have strong affinity to Con A (ca. 11% of the recovered proteins) could be fractionated within 80 min. By analysing the eluates from the column by micro two-dimensional electrophoresis, followed by blotting and Con A staining, the specificity of the column was effectively visualized. Although the protein-binding capacity of the column gradually decreased during repeated loading of serum or tissue extracts, the specificity of the column to Con A-binding proteins did not change. Serum lipoproteins have been eluted from the column with 6 M urea, suggesting that the capacity decrease is caused by the binding of lipids or lipoproteins to the column.
Manabe
T
T
Department of Chemistry, Faculty of Science, Tokyo Metropolitan University, Japan.
Higuchi
N
N
Okuyama
T
T
Mukaiyama
Y
Y
D002851:Chromatography, High Pressure Liquid; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007120:Immunochemistry; D007124:Immunoenzyme Techniques; D037102:Lectins / Q000302:isolation & purification; D011952:Receptors, Concanavalin A / Q000097:blood*
D037102:Lectins; D011952:Receptors, Concanavalin A; C027774:concanavalin A-binding glycoproteins; D003101:Collodion