Automatic Red Cell Washing Machine for Quantitative Assay of Anti‐D Concentration

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.

Section: Methodsmentioning

confidence: 99%

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

Hughes-Jones¹,

Gardner²

1973

“…Any anti-D immunoglobulin preparation of known content could be used. In this work either the WHO preparation 68/419 (provisionally assigned a value of 60 Mg/ampoule) was used, or a solution from the Bureau of Biologics/FDA (Lot 3) found to contain 300 «g anti-D immunoglobulin per ml by two analysts (six assays each on two separate days, coefficient of variation = 7%, n = 12) by the direct method of Hughes-Jones and Stevenson [2]. Different anti-D immuno globulin solutions prepared and analysed at AB Kabi were also used.…”

Section: Purification Of the Nsi Tracermentioning

confidence: 99%

“…Hughes-Jones and his colleagues introduced both the direct [2] and the indirect (or antiglobulin) method [3], [7]. The direct method is the more accurate one.…”

Section: Introductionmentioning

confidence: 99%

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

Eriksson¹,

Enell²

1976

By incorporating whole Rh(+) red cells in an agarose plate and adding a small amount of purified 125 I anti-D immunoglobulin as a tracer to standards and samples, a radioelectroimmunoassay (REIA) of anti-D immunoglobulin was possible. The results by this method agree with those obtained by the direct method. The coefficient of variation (within plate precision) is less than 9% and the reproducibility is 12% (coefficient of variation for a plasma sample assayed on six occasions during 1 month). The International anti-D serum was compared with the WHO standard preparation 68/419 on five occasions and found to contain 11; 11; 10; 9 and 11 µg per ampoule.

“…The method used for the assay of anti-D was that of Hughes-Jones and Stevenson [3], using 1251-labeIled anti-IgG globulin calibrated against «H-labelled anti-D. About one quarter of the assays were carried out manually, but the re mainder were carried out using the mechanized washing and incubating equipment described by Hughes-Jones et al [2], Only a single estimate of anti-D concentration was obtained on each sample.…”

Section: Introductionmentioning

confidence: 99%

Anti-D Immunoglobulin Preparations: the Stability of Anti-D Concentrations and the Error of the Assay of Anti-D

Hughes-Jones¹,

Hunt²,

Maycock³

et al. 1978

An analysis of the assay of 28 preparations of anti-D immunoglobulin using a radioisotope method carried out at 6-monthly intervals for 2-4.5 years showed an average fall in anti-D concentration ot 10.6% each year, with 99% confidence limits ot 6.8-14.7%. The fall in anti-D concentration after storage at 37°C for 1 month was less than 8%, the minimum change that could be detected. No significant change in physical characteristics of the immunoglobulin were detected. The error of a single estimate of anti-D by the radioisotope method (125I-labelled anti-IgG) used here was calculated to be such that the true value probably (p=0.95) lay between 66 and 150% of the estimated value.