| Autor: |
Gonzalez RM; Pacific Northwest National Laboratory, 902 Battelle Boulevard, P7-56, Richland, Washington 99354, USA., Seurynck-Servoss SL, Crowley SA, Brown M, Omenn GS, Hayes DF, Zangar RC |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of proteome research [J Proteome Res] 2008 Jun; Vol. 7 (6), pp. 2406-14. Date of Electronic Publication: 2008 Apr 19. |
| DOI: |
10.1021/pr700822t |
| Abstrakt: |
Sandwich enzyme-linked immunosorbent assay (ELISA) microarrays are emerging as a strong candidate platform for multiplex biomarker analysis because of the ELISA's ability to quantitatively measure rare proteins in complex biological fluids. Advantages of this platform are high-throughput potential, assay sensitivity and stringency, and the similarity to the standard ELISA test, which facilitates assay transfer from a research setting to a clinical laboratory. However, a major concern with the multiplexing of ELISAs is maintaining high assay specificity. In this study, we systematically determine the amount of assay interference and noise contributed by individual components of a multiplexed 24-assay system. We find that nonspecific reagent cross-reactivity problems are relatively rare. We did identify the presence of contaminant antigens in a "purified antigen". We tested the validated ELISA microarray chip using paired serum samples that had been collected from four women at a 6-month interval. This analysis demonstrated that protein levels typically vary much more between individuals than within an individual over time, a result which suggests that longitudinal studies may be useful in controlling for biomarker variability across a population. Overall, this research demonstrates the importance of a stringent screening protocol and the value of optimizing the antibody and antigen concentrations when designing chips for ELISA microarrays. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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