Electrophoretic Precipitation and Radial Diffusion Methods for Assay of Anti-D Immunoglobulin Preparations

Abstract

The measurement of anti-D concentration has been carried out by two immunoprecipitation techniques utilizing sonicated D-antigen-containing stroma suspended in agarose. One method utilizes single radial immunodiffusion; in the other method, anti-D is moved into the agarose by electrophoresis. The results obtained form both immunoprécipitation techniques were compared to those obtained by a standard 125Iantiglobulin method. The coefficient of variation of the difference between the results obtained by immunoprécipitation and those obtained from the 125I-antiglobulin method were ± 14% for the electrophoretic method and ± 16% for the radial diffusion method.

Radioelectroimmunoassay of Anti-D Immunoglobulin on Agarose Plates Containing Whole Red Cells

“…The Coombs' indirect test is only semi-quantitative. The electrophoretic precipitation assay by Hughes-Jones and Gardner [4] is sensitive enough for this type of fractions and plasmas, but they reported a large coefficient of variation (±26% for duplicates on dif ferent plates), and the preparation of stromata is difficult.…”

“…In a recent publication Hughes-Jones and Gardner [4] have reported that they have incorporated stromata of Rh-positive red cells in agarose and assayed the anti-D imunoglobulin after radial diffusion or electro phoretic precipitation.…”