Two site cross-reaction immunometric sandwich assay method

G - Physics – 01 – N

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G01N 33/573 (2006.01) G01N 33/544 (2006.01)

Patent

CA 1172958

TWO SITE CROSS-REACTION IMMUNOMETRIC SANDWICH ASSAY METHOD ABSTRACT The present invention relates generally to immunological assay techniques for determining the presence of an analyte in serum and more particularly to a two site cross-reaction immunoassay sandwich testing method which has particular application In the qualitative and quantitative determination of the level of the creatine phospho-kinase MB isoenzyme (CK-MB) in serum. Conventional immunometric techniques depend upon the immunochemical reaction between a tagged antibody which is raised to specifically react with the particular analyte of interest. However, such antibodies may not be available, or if available, then unstable. The disclosed two site-reaction immunometric sandwich assay method provides for the selection of two different antibodies each of which is specific to a different analyte but each of which will cross-react with the analyte of interest. The first antibody is reacted with the unknown sample of utilizing a solid-phase to bind the first antibody. Separation of the solid and liquid por- tions of the first reaction is accomplished and the solid portion thereof is reacted with the second antibody which is tagged. The solid portion and liquid portion of the second reaction are separ- ated and the solid portion is tested for the tag as an indication of the presence of said analyte. With particular reference to test- ing for creatine phospho-kinase-MB In human serum, the cross-reacting antibodies utilized are antibody to creatine phospho-kinase-BB and creatine phospho-kinase-MM.

380866

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