rev:February 5, 1997

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The following book on the IgG Subclass is made available by the permisssion of CLB Reagents (Netherlands). It is for your personal use and may not be reproduced without the permission of CLB Reagents. This material is for information use only and should not be construed as medical advice. We are not responsible for any misprints or error or ommissions. Certain charts ,graphs and table have been ommitted or changed in style (not content) to fit this web site. Please use this information as part of your discussion with your medical professional.

5    Assays for the determination of IgG subclass levels

Different methods are in use for the quantitative determination of IgG subclasses. For most methods, The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service (CLB) is a manufacturer of special reagents and kits:

5.1     Radial Immunodiffusion (RID)

RID (Mancini) is a classical diagnostic method to determine IgG subclasses. This reliable assay is widely used and easy to perform. However, when large numbers of samples have to be processed the relatively long incubation times will be a disadvantage.

Brief outline of the method:

The RID assay is performed in (ready-for-use) agar plates,containing the specific anti-IgG subclass antibodies (figure 7). Test samples, standard-and control sera are prepared and added to the plates. After 48-64 hours incubation at room temperature the diameters of the immunoprecipitation rings are measured. The IgG subclass concentrations in the test samples may be quantified in two ways:
a) Calibration curve method: ring diameters and concentrations of the standards are plotted and the values of the test sample are determined by interpolation.

b) Tabular method: ring diameters of the calibration curve are listed and the values of the test sample are read from a table. It is not necessary to make a calibration curve. The control serum is assayed to check the validity of the calibration curves and also the accuracy of the IgG subclass quantification, when using the table.

-reference list

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