Label-Free Screening of Protein Binding to Small Molecule Compound Microarrays with A High-Throughput Optical Scanning Microscope

2009 ◽  
Author(s):  
J. P. Landry ◽  
Y. Y. Fei ◽  
Y. S. Sun ◽  
J. T. Luo ◽  
K. S. Lam ◽  
...  
Keyword(s):  
Protein Binding ◽  
Small Molecule ◽  
High Throughput ◽  
Label Free ◽  
Scanning Microscope ◽  
Optical Scanning ◽  
Small Molecule Compound

scholarly journals A novel high-throughput scanning microscope for label-free detection of protein and small-molecule chemical microarrays

2008 ◽  
Vol 79 (1) ◽  
pp. 013708 ◽  
Author(s):  
Y. Y. Fei ◽  
J. P. Landry ◽  
Y. S. Sun ◽  
X. D. Zhu ◽  
J. T. Luo ◽  
...  
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
Label Free ◽  
Scanning Microscope ◽  
Label Free Detection

scholarly journals Label-free profiling of DNA aptamer-small molecule binding using T5 exonuclease

2020 ◽  
Vol 48 (20) ◽  
pp. e120-e120 ◽  
Author(s):  
Obtin Alkhamis ◽  
Weijuan Yang ◽  
Rifat Farhana ◽  
Haixiang Yu ◽  
Yi Xiao
Keyword(s):  
Ligand Binding ◽  
Small Molecule ◽  
High Throughput ◽  
Dna Aptamer ◽  
Label Free ◽  
General Applicability ◽  
Gold Standard Method ◽  
Characterization Methods ◽  
Increased Risk

Abstract In vitro aptamer isolation methods can yield hundreds of potential candidates, but selecting the optimal aptamer for a given application is challenging and laborious. Existing aptamer characterization methods either entail low-throughput analysis with sophisticated instrumentation, or offer the potential for higher throughput at the cost of providing a relatively increased risk of false-positive or -negative results. Here, we describe a novel method for accurately and sensitively evaluating the binding between DNA aptamers and small-molecule ligands in a high-throughput format without any aptamer engineering or labeling requirements. This approach is based on our new finding that ligand binding inhibits aptamer digestion by T5 exonuclease, where the extent of this inhibition correlates closely with the strength of aptamer-ligand binding. Our assay enables accurate and efficient screening of the ligand-binding profiles of individual aptamers, as well as the identification of the best target binders from a batch of aptamer candidates, independent of the ligands in question or the aptamer sequence and structure. We demonstrate the general applicability of this assay with a total of 106 aptamer-ligand pairs and validate these results with a gold-standard method. We expect that our assay can be readily expanded to characterize small-molecule-binding aptamers in an automated, high-throughput fashion.

scholarly journals A fluorescence-based high throughput assay for the determination of small molecule−human serum albumin protein binding

2014 ◽  
Vol 406 (7) ◽  
pp. 1867-1875 ◽  
Author(s):  
Megan M. McCallum ◽  
Alan J. Pawlak ◽  
William R. Shadrick ◽  
Anton Simeonov ◽  
Ajit Jadhav ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Protein Binding ◽  
Human Serum ◽  
Small Molecule ◽  
High Throughput ◽  
High Throughput Assay
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