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High-throughput multiplex protein analysis for limited sample size on a single slide

S. Rozhok, K. Ouyang, D. Dixon, M. Nelson
NanoInk, Inc., US

Keywords: microarray, cytokines


To meet the challenges of new genomic and proteomic applications, rapid and sensitive analysis of large numbers of interactions in parallel will be required. High-density microarrays are ideally suited for parallel multiplex screening of thousands of interactions with minimum use of materials. In this study, we report on high throughput screening of 10 different cytokines using both 48 and 96 well formats on a single slide. Multiple arrays of cytokine antibodies were printed on a single microarray slide utilizing Dip Pen Nanolithography® (DPN®). DPN allows parallel printing of hundreds of molecules over an area that is 100 times smaller than that of conventional microarray formats. Smaller feature sizes require significantly smaller amounts of synthesized and labeled materials, which is especially pertinent to drug targets that are expressed in vanishingly small quantities. The unique design of this high throughput format does not require gaskets, allowing reaction volumes of only a few microliters. To validate this new approach both conventional and high throughput formats were run in parallel and the results were compared. The performance of these high throughput formats meets or exceeds the performance of conventional assays, while requiring significantly smaller volumes of sample and reagents. Additionally, the gasket-less approach offers up to 384 individual reaction wells and sample sizes as low as 1 microliter affording the capability for parallel quantitative measurements on a scale not previously available.
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