期刊
JOURNAL OF PROTEOME RESEARCH
卷 7, 期 6, 页码 2406-2414出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr700822t
关键词
antibody; ELISA; microarray; cross-reactivity; longitudinal; serum
资金
- NCI NIH HHS [CA117378] Funding Source: Medline
- NIBIB NIH HHS [EB006177] Funding Source: Medline
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.
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