Slide immunoenzymatic assay for immunoglobulin isotype (SIA-Ig).
A simple method is described for rapid determination of immunoglobulin class and subclass in an assortment of samples based on the slide immunoenzymatic assay (SIA-Ig). Each circle on a multi-circle glass slide is coated with anti-immunoglobulin class or subclass antibody. For each isotype to be assayed a circle is coated with its specific anti-isotype. The coated circles are incubated with sample containing the immunoglobulin(s) of unknown isotype and are then washed. The slide is then incubated with enzyme-labeled anti-immunoglobulin and washed again. Finally, enzyme substrate is deposited onto the circles. Color appears within a few minutes only on the circle where the unknown was bound specifically by its corresponding anti-isotype antibody. The method reveals correctly the isotype of the constituents of complex mixtures, such as serum, as well as that of the only component of samples containing a single molecular species of immunoglobulin (e.g., monoclonal antibodies). The method is simple, reliable, gives results in less than 1 h and is adequately sensitive for a wide range of practical applications.
D000469:Alkaline Phosphatase; D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D005779:Immunodiffusion; D007124:Immunoenzyme Techniques; D007126:Immunoglobulin Allotypes / Q000032:analysis / Q000145:classification*; D007074:Immunoglobulin G / Q000145:classification / Q000276:immunology; D051379:Mice; D011817:Rabbits; D013997:Time Factors; D001616:beta-Galactosidase
D000888:Antibodies, Anti-Idiotypic; D007126:Immunoglobulin Allotypes; D007074:Immunoglobulin G; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase