ABSTRACT
Human immunoglobulins (IgG) are produced on a multi-ton scale for therapeutic applications. There is presently no available method to manufacture IgG preparations enriched with immunoglobulins from the IgG2 subclass although they might be useful for therapeutic purposes. By frontal chromatography, we have screened 69 immobilized dyes, among which, six display a different affinity for IgG2 and other subclasses. One (Procion Yellow HE-4R) was studied further. The screening of various mobile phase conditions allowed us to devise a procedure to prepare IgG2 enriched IgG solutions: The cumulative yield for IgG2 was 43% and IgG2/total IgG ratio in the final product was 67%.
Subject(s)
Coloring Agents/chemistry , Immunoglobulin G/isolation & purification , Triazines/chemistry , Affinity Labels , Chemical Phenomena , Chemistry, Physical , Chromatography , HumansABSTRACT
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.
