Detection of oligonucleotide-ligand complexes by ESI-MS (DOLCE-MS) as a component of high throughput screening.
With the advent of combinatorial chemistry and high throughput screening, a major bottleneck in the pharmaceutical industry has changed from quickly finding active compounds to limiting them to a manageable number for proper follow-up. With hundreds to thousands of active compounds identified by a multitude of biological screens, there need to be rapid and unambiguous methods for eliminating false positive, toxic, or otherwise difficult compounds from further scrutiny. We have used electrospray ionization mass spectrometry as a rapid screening method to identify compounds from viral screens that yield a positive assay response by interaction with DNA rather than inhibiting the target enzyme. Both the sample preparation and data acquisition have been automated, allowing the screening of all hits from relevant biological screens (up to 1,000/week). The assay was validated using several known DNA intercalators and minor groove binders. These "standards" and many but not all of our "active compounds" were shown to form noncovalent complexes with a variety of different DNA:DNA and DNA:RNA duplexes.
Greig
M J
MJ
Pfizer Global R&D, San Diego, CA 92121, USA. michael.greig@agouron.com
Robinson
J M
JM
D001483:Base Sequence; D020650:Combinatorial Chemistry Techniques; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods* / Q000592:standards; D007364:Intercalating Agents; D008024:Ligands; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis; D009841:Oligonucleotides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D012313:RNA / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards
D007364:Intercalating Agents; D008024:Ligands; D009692:Nucleic Acid Heteroduplexes; D009841:Oligonucleotides; D012313:RNA; D004247:DNA