Monoclonal immunoglobulins: affinity blotting for low concentrations in serum.
I describe a simple, economical technique for identifying low concentrations of monoclonal immunoglobulins in the presence of excessive amounts of immunoglobulins of other classes. The technique involves binding of specific antibody to nitrocellulose, separating proteins by isoelectric focusing or zone electrophoresis in agarose gels, using capillary transfer to bind proteins to the nitrocellulose via their antibody affinity, and then detecting transferred proteins with enzyme-labeled antibody. A monoclonal immunoglobulin can be completely characterized in 2 h. No expensive equipment is required. Affinity blotting is about 10-fold as sensitive as native blotting, 100-fold as sensitive as silver staining.
McLachlan
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Pathology Department, Peter MacCallum Cancer Institute, Melbourne, Australia.
D000911:Antibodies, Monoclonal / Q000032:analysis*; D003101:Collodion; D004587:Electrophoresis, Agar Gel; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007072:Immunoglobulin D / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D007136:Immunoglobulins / Q000032:analysis*; D007525:Isoelectric Focusing
D000911:Antibodies, Monoclonal; D007072:Immunoglobulin D; D007075:Immunoglobulin M; D007136:Immunoglobulins; D003101:Collodion