Chromatography of human immunoglobulin G on immobilized drimarene rubine R/K-5BL. Study of mild, efficient elution procedures.
A range of substances were screened to find eluents for human immunoglobulin G (IgG) which are retained with a strong affinity by immobilized Drimarene Rubine R/K-5BL. The strong affinity of IgG for the dye is partly due to the presence of copper in the dye. This was suggested by the effect of substances able to make coordination bonds with metals that elute the IgG and also the effect of metal stripping from the immobilized dye. Several mobile phase conditions were found that allowed desorption of retained IgG on immobilized Drimarene Rubine R/K-5BL without using a protein denaturant. A procedure was also devised for separating IgG2 from other IgG subclasses using chromatography on immobilized Rubine R/K-5BL and column development with an AMP gradient.
Cochet
S
S
INSERM U 76, Paris, France.
Hasnaoui
M H
MH
Debbia
M
M
Kroviarski
Y
Y
Lambin
P
P
Cartron
J P
JP
Bertrand
O
O
D000588:Amines / Q000737:chemistry; D000596:Amino Acids / Q000737:chemistry; D002021:Buffers; D002847:Chromatography, Agarose / Q000379:methods; D004396:Coloring Agents; D004492:Edetic Acid / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007093:Imidazoles / Q000737:chemistry; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008670:Metals / Q000737:chemistry; D009705:Nucleosides / Q000737:chemistry; D009942:Organometallic Compounds; D010710:Phosphates / Q000737:chemistry; D010920:Placenta / Q000737:chemistry / Q000276:immunology; D011247:Pregnancy; D011506:Proteins / Q000302:isolation & purification; D012394:Rosaniline Dyes; D012685:Sepharose / Q000031:analogs & derivatives
D000588:Amines; D000596:Amino Acids; D002021:Buffers; D004396:Coloring Agents; C087073:Drimarene rubine R-K-5BL; D007093:Imidazoles; D007074:Immunoglobulin G; D008670:Metals; D009705:Nucleosides; D009942:Organometallic Compounds; D010710:Phosphates; D011506:Proteins; D012394:Rosaniline Dyes; C035771:Sepharose CL 4B; D012685:Sepharose; D004492:Edetic Acid; C004692:coomassie Brilliant Blue