Determination of Proteins in Biological Fluids by Electroimmunodiffusion and Two-Directional Immunoelectrophoresis

[摘要] We have used electroimmunodiffusion and two-directional immunoelectrophoresis to measure proteins in biological fluids. Electroimmunodiffusion is based on the electrophoresis of antigens in antibody-containing agarose under standardized conditions. Pre-albumin, albumin, α1antitrypsin, α2-macroglobulin, IgG, IgA, IgM, IgD, and other proteins have been determined in our laboratory with a precision of ±5%. The lower limit for quantitation is 5 µg/ml. For immunoglobulin quantitation, treatment of the samples with potassium cyanate (carbamylation) before electrophoresis improves sensitivity and accuracy. Two-directional immunoelectrophoresis is based on electrophoretic separation of proteins in agarose or other high-resolution media followed by electrophoresis at right angles to the first direction, in a gel in which an appropriate antiserum has been incorporated. Qualitative and semiquantitative changes in proteins in abnormal specimens can be assessed. The two techniques combined provide a very sensitive tool for the evaluation of protein changes in biologic fluids. A simple apparatus has been designed in which a number of specimens can be analyzed simultaneously by either method.

[学科分类] 过敏症与临床免疫学