Sandwich-type, antibody microarrays for the detection and quantification of cardiovascular risk markers

A sandwich-type antibody microarray was developed for the detection and quantitation of cardiovascular risk markers including C-reactive protein, TNF-α, serum amyloid-A, fatty acid binding protein. In this assay, antibodies to cardiovascular risk marker protein antigens were immobilized on super-epoxy coated glass slides as microarrays. These microarrays were then probed with pure antigens or human sera followed by biotinylated detection antibodies. Biotinylated antibodies were scanned and quantitated for flourescence after reaction with flourescent streptavidin. Results indicate that baseline and elevated levels of C-reactive protein, myoglobin, TNF-α, serum amyloid-A can be detected and quantified. This study is the first study of its kind, utilizing sandwich-type arrays for the detection and quantification of multiple-cardiovascular risk markers in human serum. We have also presented, in this study, that anti-fatty-acid binding protein antibodies, used as capture molecules, cross reacts with all detection antibodies on the microarray platform, generating noise signals, interfering the detection and quantification of markers. Finally, we have compared the results of our study with the performance of commercially available ELISA test-kits that presenting the advantages such as being able to detect and quantify multiple markers at once and also better dynamic ranges over some of the commercial ELISA test-kits.