Multiplex bead array assays: performance evaluation and comparison of sensitivity to ELISA.
The measurement of soluble cytokines and other analytes in serum and plasma is becoming increasingly important in the study and management of many diseases. As a result, there is a growing demand for rapid, precise, and cost-effective measurement of such analytes in both clinical and research laboratories. Multiplex bead array assays provide quantitative measurement of large numbers of analytes using an automated 96-well plate format. Enzyme-linked immunosorbent assay (ELISAs) have long been the standard for quantitative analysis of cytokines and other biomarkers, but are not well suited for high throughput multiplex analyses. However, prior to replacement of ELISA assays with multiplex bead array assays, there is a need to know how comparable these two methods are for quantitative analyses. A number of published studies have compared these two methods and it is apparent that certain elements of these assays, such as the clones of monoclonal antibodies used for detection and reporting, are pivotal in obtaining similar results from both assays. By careful consideration of these variables, it should be possible to utilize multiplex bead array assays in lieu of ELISAs for studies requiring high throughput analysis of numerous analytes.
National Heart, Lung, and Blood Institute, NIH Bethesda, MD 20892, USA.
Mohamed F
MF
Elshal
J Philip
JP
McCoy
D000911:Antibodies, Monoclonal / Q000737:chemistry; D001681:Biological Assay; D015415:Biomarkers; D003362:Cost-Benefit Analysis; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008027:Light; D015999:Multivariate Analysis; D040081:Protein Array Analysis / Q000379:methods*; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D016207:Cytokines; D008024:Ligands