High-throughput screening in two dimensions: Binding intensity and off-rate on a peptide microarray

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• 作者：Matthew P. Greving ; Paul E. Belcher ; Conor D. Cox ; Douglas Daniel ; Chris W. Diehnelt ; Neal W. Woodbury
• 刊名：Analytical Biochemistry
• 出版年：2010
• 出版时间：1 July 2010
• 年：2010
• 卷：402
• 期：1
• 页码：93-95
• 全文大小：223 K

文摘

We report a high-throughput two-dimensional microarray-based screen, incorporating both target binding intensity and off-rate, which can be used to analyze thousands of compounds in a single binding assay. Relative binding intensities and time-resolved dissociation are measured for labeled tumor necrosis factor alpha (TNF-α) bound to a peptide microarray. The time-resolved dissociation is fitted to a one-component exponential decay model, from which relative dissociation rates are determined for all peptides with binding intensities above background. We show that most peptides with the slowest off-rates on the microarray also have the slowest off-rates when measured by surface plasmon resonance (SPR).