Methods for screening compound libraries
OwnerOle Hindsgaul, David C. Schriemerinventors
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Abstract
Disclosed are methods for screening compound libraries using frontal chromatography in combination with mass spectrometry to identify and rank those members of the library that bind to a target receptor. Methods are also disclosed which permit a compound library to be rapidly screened to determine if any members of the library have a higher affinity for the target receptor relative to a pre-selected indicator compound.
Claims43
A method for screening a compound library to determine the relative or absolute affinity of a plurality of putative ligands to a target receptor, which method comprises:
- (a) providing a compound library comprising a plurality of putative ligands,
- (b) continuously applying the compound library to a column comprising a target receptor selected from the group consisting of peptides, proteins, glycoproteins, glycosaminoglycans, proteoglycans, integrins, enzymes, lectins, selectins, cell-adhesion molecules, toxins, transport proteins, hormones, antibodies, cadherins, DNA, DNA fragments, RNA and RNA fragments, optionally immobilized on a solid phase support under frontal chromatography conditions whereby the target receptor is continuously contacted with the compound library to provide an effluent;
- (c) continuously or intermittently applying the effluent to a mass spectrometer to provide mass spectra of the constituent putative ligands present in the effluent;
- (d) evaluating the mass spectra to determine a break through time for each of the putative ligands;
- and
- (e) determining an affinity to the target receptor for a putative ligand in each library relative to another putative ligand in the same library by comparing the break through time on the column for the putative ligand relative to the other putative ligand in the same library.
A method for screening a plurality of compound libraries to determine the relative or absolute affinity of a putative ligands in each library to a target receptor, which method comprises:
- (a) providing a plurality of compound libraries, each library comprising a plurality of putative ligands,
- (b) continuously applying each compound library to a separate column comprising a target receptor independently selected from the group consisting of peptides, proteins, glycoproteins, glycosaminoglycans, proteoglycans, integrins, enzymes, lectins, selectins, cell-adhesion molecules, toxins, transport proteins, hormones, antibodies, cadherins, DNA, DNA fragments, RNA and RNA fragments optionally immobilized on a solid phase support under frontal chromatography conditions whereby the target receptor is continuously contacted with the compound library to provide an effluent from each column;
- (c) intermittently applying the effluent from each column to a mass spectrometer to provide mass spectra of the constituent putative ligands present in the effluent;
- (d) evaluating the mass spectra to determine a break through time for each of the putative ligands in each compound library;
- and
- (e) determining a relative affinity for each of the putative ligands in each library to the target receptor by comparing the break through time on the column for each of the putative ligands in each library relative to the other putative ligands in the same library.
The method of wherein the target receptor is a DNA fragment.
Dependencies are inferred from the claim text.