Screening small-molecule compound microarrays for protein ligands without fluorescence labeling with a high-throughput scanning microscope.
We describe a high-throughput scanning optical microscope for detecting small-molecule compound microarrays on functionalized glass slides. It is based on measurements of oblique-incidence reflectivity difference and employs a combination of a y-scan galvometer mirror and an x-scan translation stage with an effective field of view of 2 cm x 4 cm. Such a field of view can accommodate a printed small-molecule compound microarray with as many as 10,000 to 20,000 targets. The scanning microscope is capable of measuring kinetics as well as endpoints of protein-ligand reactions simultaneously. We present the experimental results on solution-phase protein reactions with small-molecule compound microarrays synthesized from one-bead, one-compound combinatorial chemistry and immobilized on a streptavidin-functionalized glass slide.
Fei
Yiyan
Y
University of California at Davis, Department of Physics, Davis, California 95616, USA.
Landry
James P
JP
Sun
Yungshin
Y
Zhu
Xiangdong
X
Wang
Xiaobing
X
Luo
Juntao
J
Wu
Chun-Yi
CY
Lam
Kit S
KS
D000818:Animals; D001710:Biotin / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D019169:Jurkat Cells; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D040081:Protein Array Analysis / Q000379:methods*; D054852:Small Molecule Libraries / Q000378:metabolism; D019809:Streptavidin / Q000378:metabolism
D007074:Immunoglobulin G; D054852:Small Molecule Libraries; D001710:Biotin; D019809:Streptavidin