A sensitive double-diffusion microassay suitable for the detection of idiotype-antiidiotype precipitates.
Using agarose gel coated on GelBond film sheets and using Coomassie blue stain followed by silver stain, a sensitive double-diffusion microassay has been developed for detecting small amounts of precipitate forming during idiotype-antiidiotype reactions. The sensitivity of the method is 10-100 times greater than classical immunodiffusion tests. Other advantages include the need for minimal amounts of unconcentrated sample, the absence of radioactive or toxic substrates, no interference due to a second or third antibody coat such as are used in immunoenzymatic techniques, and the possibility of a direct evaluation of qualitative data such as identity, cross-reactivity or non-identity. As little as 40 ng antibody could be detected, corresponding to an antibody concentration of 8 micrograms/ml, making the microassay useful for rapid screening of idiotype-antiidiotype precipitates during routine analysis of hybridoma supernatants.
Rochu
D
D
Department of Protein Immunochemistry, Institut National de Transfusion Sanguine, Paris, France.
Crespeau
H
H
Fine
A
A
Fine
J M
JM
D000017:ABO Blood-Group System / Q000276:immunology; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D003429:Cross Reactions; D004306:Dose-Response Relationship, Immunologic; D005779:Immunodiffusion / Q000379:methods*; D007130:Immunoglobulin Idiotypes / Q000276:immunology*; D007158:Immunologic Techniques; D051379:Mice; D011233:Precipitin Tests / Q000379:methods*; D011817:Rabbits
D000017:ABO Blood-Group System; D000888:Antibodies, Anti-Idiotypic; D007130:Immunoglobulin Idiotypes