Abstract
This chapter discusses the isolation of soluble immune complexes (IC) from human serum, with combined use of polyethylene glycol (PEG) precipitation, gel filtration, and affinity chromatography on protein. The most popular approach to the isolation of ICs has been the use of immobilized staphylococcal protein A as a substrate. PEG is known to precipitate antigen-antibody complexes more readily than free antigen or free antibody. It is found that at the concentration used for first step, IgM, IgG, a2-macroglobulin, and C4 are also precipitated even in their native forms. Additional steps are needed to separate the ICs from these proteins. Gel filtration on any of the several types of gel can be used effectively to separate the high molecular-weight fractions from monomeric IgG. Double immunodiffusion analysis showed that pool III contained IgG only, indicating that this patient had a mixture of atypical anti-immunoglobulin antibodies, probably including IgM anti-IgA and IgG anti-IgG. The results for the ICs formed in vitro indicate that gel filtration on sepharose G-50, under dissociating conditions, is an effective approach to the dissociation of insulin-anti-insulin IC.
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