High-throughput screening in two dimensions: binding intensity and off-rate on a peptide microarray.
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-alpha) 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).
Greving
Matthew P
MP
The Biodesign Institute at Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ 85287-5201, USA.
Belcher
Paul E
PE
Cox
Conor D
CD
Daniel
Douglas
D
Diehnelt
Chris W
CW
Woodbury
Neal W
NW
D000595:Amino Acid Sequence; D057166:High-Throughput Screening Assays / Q000379:methods; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D020349:Surface Plasmon Resonance / Q000379:methods*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D019151:Peptide Library; D010455:Peptides; D014409:Tumor Necrosis Factor-alpha