Microfluorometric evaluation of the specificity of fluorescent antisera against mouse immunoglobulins with the defined antigen substrate spheres (DASS) system
article
Highly purified MOPC 21 IgG1, MOPC 173 IgG(2a), MOPC 195 IgG(2b), MOPC 104 E IgM, and MOPC 315 IgA paraproteins, heterogeneous mouse IgG, Fab and Fc fragments of heterogeneous IgG were prepared and coupled to Sepharose beads. These beads were then used as artificial substrates to test the specificity of fluorescent antisera against mouse immunoglobulins by microfluorometry. By comparing the visual evaluation of stained plasma cells and measurements on beads, the highest permissible percentage impurity in a conjugate was determined. It was 14% for a fluorescein iso thiocyanate and 6% for a tetramethyl rhodamine iso thiocyanate conjugate. By application of these criteria, 1 out of 7 tested commercial antisera and 6 out of 8 conjugates prepared in this laboratory proved to be satisfactory. The most common impurities were anti light chain antibodies, as revealed by their reaction with Fab. With the bead system, a good impression of the specificity of an antiserum can be obtained. It gives, however, only approximate information on whether the conjugate will cause a high background staining in the biological specimen.
Chemicals/CAS: Antibodies, Anti-Idiotypic; Antigens; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Sepharose, 9012-36-6
Chemicals/CAS: Antibodies, Anti-Idiotypic; Antigens; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Sepharose, 9012-36-6
Topics
TNO Identifier
228008
ISSN
00221759
Source
Journal of Immunological Methods, 10(4), pp. 337-355.
Pages
337-355
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