Claims
- 1. An apparatus for screening a compound library to determine the relative or absolute affinity of a plurality of putative ligands to a target receptor or a plurality of target receptors, which apparatus comprises:
(a) a column comprising a target receptor or a plurality of target receptors, each target receptor optionally attached to a solid phase support, and having a inflow end and an outflow end, wherein said column is capable of having a compound library comprising a plurality of putative ligands applied thereto under frontal chromatography conditions to produce an effluent from the outflow end of the column; (b) a first reservoir connected to the inflow end of said column for applying the compound library to the column; (c) a mass spectrometer connected to the outflow end of said column for continuously or intermittently analyzing the effluent from the column.
- 2. The apparatus of claim 1, wherein said apparatus further comprises:
(d) a second reservoir connected to the inflow end of the column for applying either (i) a mixture comprising the compound library, at least one void marker compound and an indicator compound or a plurality of indicators compounds, (ii) at least one void marker compound and an indicator compound or a plurality of indicator compounds, or (iii) a buffer solution to the column.
- 3. The apparatus of claim 1, wherein said apparatus further comprises:
(e) a third reservoir connected to the outflow end of the column for supplying a supplemental diluent to the effluent before analysis by the mass spectrometer.
- 4. The apparatus of claim 1, wherein the column has an internal diameter ranging from about 10 μm to about 4.6 mm.
- 5. The apparatus of claim 4, wherein the column has an internal diameter of from about 100 μm to about 250 μm.
- 6. The apparatus of claim 1, wherein the column has a length of from about 1 cm to about 30 cm.
- 7. The apparatus of claim 1, wherein the column has a length of from about 2 cm to about 20 cm.
- 8. The apparatus of claim 1, wherein each target receptor is independently selected from the group consisting of proteins, glycoproteins, glycosaminoglycans, proteoglycans, integrins, enzymes, lectins, selecting, cell-adhesion molecules, toxins, bacterial pili, transport proteins, receptors involved in signal transduction or hormone-binding, hormones, antibodies, major histocompatability complexes, immunoglobulin superfamilies, cadherins, DNA or DNA fragments, RNA and RNA fragments, whole cells, cell fragments, tissues, bacteria, fungi, viruses, parasites, preons, and synthetic analogs or derivatives thereof.
- 9. The apparatus of claim 1, wherein the target receptor is bound to a solid phase support.
- 10. The apparatus of claim 9, wherein the target receptor is covalently bound to the solid phase support or bound via biotin-avidin or biotin-streptavidin binding.
- 11. The apparatus of claim 9, wherein the solid phase support is selected from the group consisting of polymeric beads, polymeric gels, glass beads, silica chips, silica capillaries, agarose, diatomaceous earths and pulp.
- 12. The apparatus of claim 1, wherein the column contains from about 1 fmol to about 10 nmol of target receptor active sites.
- 13. The apparatus of claim 1, wherein the mass spectrometer is an electrospray mass spectrometer.
- 14. An apparatus for screening a plurality of compound libraries to determine the relative or absolute affinity of a plurality of putative ligands in each library to a target receptor or a plurality of target receptors, which apparatus comprises:
(a) a plurality of columns each column comprising a target receptor or a plurality of target receptors, each target receptor optionally attached to a solid phase support, and each column having a inflow end and an outflow end, wherein each of said columns is capable of independently having a compound library comprising a plurality of putative ligands applied thereto under frontal chromatography conditions to produce an effluent from the outflow end of the column; (b) a plurality of first reservoirs each connected to the inflow end of one of the columns for applying a compound library to the columns; (c) a mass spectrometer connected to the outflow end of each of said columns for intermittently analyzing the effluent from each of the column.
- 15. The apparatus of claim 14, wherein said apparatus further comprises:
(d) a plurality of second reservoirs each connected to the inflow end of one of the columns for applying either (i) a mixture comprising the compound library, at least one void marker compound and an indicator compound or a plurality of indicator compounds, (ii) at least one void marker compound and an indicator compound or a plurality of indicator compounds, or (iii) a buffer solution to the column.
- 16. The apparatus of claim 14, wherein said apparatus further comprises:
(e) a third reservoir connected to the outflow end of each of the columns for supplying a supplemental diluent to the effluent from each column before analysis by the mass spectrometer.
- 17. The apparatus of claim 14, wherein said apparatus comprises from 2 to about 100 columns.
- 18. The apparatus of claim 17, wherein said apparatus comprises from 3 to about 50 columns.
- 19. The apparatus of claim 18, wherein said apparatus comprises from 5 to about 10 columns.
- 20. The apparatus of claim 14, wherein each column is intermittently monitored for a period of about 0.5 seconds to about 10 seconds before switching to the next column.
- 21. The apparatus of claim 20, wherein each column is intermittently monitored for about 1 second to about 5 seconds before switching to the next column.
- 22. The apparatus of claim 14, wherein the column has an internal diameter ranging from about 10 μm to about 4.6 mm.
- 23. The apparatus of claim 22, wherein the column has an internal diameter of from about 100 μm to about 250 μm.
- 24. The apparatus of claim 14, wherein the column has a length of from about 1 cm to about 30 cm.
- 25. The apparatus of claim 14, wherein the column has a length of from about 2 cm to about 20 cm.
- 26. The apparatus of claim 14, wherein each target receptor is in dependently selected from the group consisting of proteins, glycoproteins, glycosaminoglycans, proteoglycans, integrins, enzymes, lectins, selectins, cell-adhesion molecules, toxins, bacterial pili, transport proteins, receptors involved in signal transduction or hormone-binding, hormones, antibodies, major histocompatability complexes, immunoglobulin superfamilies, cadherins, DNA or DNA fragments, RNA and RNA fragments, whole cells, cell fragments, tissues, bacteria, fungi, viruses, parasites, preons, and synthetic analogs or derivatives thereof.
- 27. The apparatus of claim 14, wherein each target receptor is bound to a solid phase support.
- 28. The apparatus of claim 27, wherein each target receptor is covalently bound to the solid phase support or bound via biotin-avidin or biotin-streptavidin binding.
- 29. The apparatus of claim 27, wherein the solid phase support is selected from the group consisting of polymeric beads, polymeric gels, glass beads, silica chips, silica capillaries, agarose, diatomaceous earths and pulp.
- 30. The apparatus of claim 14, wherein the column contains from about 1 fmol to about 10 nmol of target receptor active sites.
- 31. The apparatus of claim 14, wherein the mass spectrometer is an electrospray mass spectrometer.
- 32. An apparatus for screening a target receptor or a plurality of target receptors to determine the relative affinity of the receptor or receptors to an immobilized ligand or ligands relative to an indicator compound or a plurality of indicator compounds, which apparatus comprises:
(a) a column comprising a ligand or a plurality of ligands wherein each ligand is bound to a solid phase support, said column having a inflow end and an outflow end and further wherein said column is capable of having a target receptor or a plurality of target receptors applied thereto under frontal chromatography conditions to produce an effluent from the outflow end of the column; (b) a first reservoir connected to the inflow end of said column for applying the target receptor or receptors to the column; (c) a second reservoir connected to the inflow end of the column for applying either (i) a mixture comprising the target receptor or receptors, at least one void marker compound and an indicator compound or a plurality of indicators compounds, (ii) at least one void marker compound and an indicator compound or a plurality of indicator compounds, or (iii) a buffer solution to the column. (d) a mass spectrometer connected to the outflow end of said column for continuously or intermittently analyzing the effluent from the column.
- 33. The apparatus of claim 32, wherein said apparatus further comprises:
(e) a third reservoir connected to the outflow end of the column for supplying a supplemental diluent to the effluent before analysis by the mass spectrometer.
- 34. The apparatus of claim 32, wherein the column has an internal diameter ranging from about 10 μm to about 4.6 mm.
- 35. The apparatus of claim 34, wherein the column has an internal diameter of from about 100 μm to about 250 μm.
- 36. The apparatus of claim 32, wherein the column has a length of from about 1 cm to about 30 cm.
- 37. The apparatus of claim 32, wherein the column has a length of from about 2 cm to about 20 cm.
- 38. The apparatus of claim 32, wherein each ligand is selected from the group consisting of carbohydrates, monosaccharides, oligosaccharides, polysaccharides, amino acids, peptides, oligopeptides, polypeptides, proteins, nucleosides, nucleotides, oligonucleotides, polynucleotides, lipids, retinoids, steroids, glycopeptides, glycoproteins, glycolipids, proteoglycans, and synthetic analogs or derivatives thereof.
- 39. The apparatus of claim 32, wherein each ligand is selected from the group consisting of synthetic small molecule organic compounds.
- 40. An apparatus for screening a plurality of target receptors to determine the relative affinity of the receptors to an immobilized ligand or ligands relative to an indicator compound or a plurality of indicator compounds, which apparatus comprises:
(a) a plurality of columns each column comprising a ligand or a plurality of ligands wherein each ligand is bound to a solid phase support, and each column having a inflow end and an outflow end, wherein each of said columns is capable of independently having a target receptor or a plurality of target receptors applied thereto under frontal chromatography conditions to produce an effluent from the outflow end of the column; (b) a plurality of first reservoirs each connected to the inflow end of one of the columns for applying a target receptor or a plurality of target receptors to the columns; (c) a plurality of second reservoirs each connected to the inflow end of one of the columns for applying either (i) a mixture comprising the target receptor or plurality of target receptors, at least one void marker compound and an indicator compound or a plurality of indicator compounds, (ii) at least one void marker compound and an indicator compound or a plurality of indicator compounds, or (iii) a buffer solution to the column. (d) a mass spectrometer connected to the outflow end of each of said columns for intermittently analyzing the effluent from each of the column.
- 41. The apparatus of claim 40, wherein said apparatus further comprises:
(e) a third reservoir connected to the outflow end of each of the columns for supplying a supplemental diluent to the effluent from each column before analysis by the mass spectrometer.
- 42. The apparatus of claim 40, wherein said apparatus comprises from 2 to about 100 columns.
- 43. The apparatus of claim 42, wherein said apparatus comprises from 3 to about 50 columns.
- 44. The apparatus of claim 43, wherein said apparatus comprises from 5 to about 10 columns.
- 45. The apparatus of claim 40, wherein each column is intermittently monitored for a period of about 0.5 seconds to about 10 seconds before switching to the next column.
- 46. The apparatus of claim 45, wherein each column is intermittently monitored for about 1 second to about 5 seconds before switching to the next column.
- 47. The apparatus of claim 40, wherein the column has an internal diameter ranging from about 10 μm to about 4.6 mm.
- 48. The apparatus of claim 47, wherein the column has an internal diameter of from about 100 μm to about 250 μm.
- 49. The apparatus of claim 40, wherein the column has a length of from about 1 cm to about 30 cm.
- 50. The apparatus of claim 40, wherein the column has a length of from about 2 cm to about 20 cm.
- 51. The apparatus of claim 40, wherein each ligand is selected from the group consisting of carbohydrates, monosaccharides, oligosaccharides, polysaccharides, amino acids, peptides, oligopeptides, polypeptides, proteins, nucleosides, nucleotides, oligonucleotides, polynucleotides, lipids, retinoids, steroids, glycopeptides, glycoproteins, glycolipids, proteoglycans, and synthetic analogs or derivatives thereof.
- 52. The apparatus of claim 40, wherein each ligand is selected from the group consisting of synthetic small molecule organic compounds.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of U.S. Ser. No. 09/069,890, filed Apr. 29, 1998, which application claims the benefit of U.S. Provisional Application No. 60/079,622, filed Mar. 27, 1998. Each of these applications are incorporated herein by reference in their entirety.
Provisional Applications (1)
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Number |
Date |
Country |
|
60079622 |
Mar 1998 |
US |
Divisions (1)
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Number |
Date |
Country |
Parent |
09276443 |
Mar 1999 |
US |
Child |
10093484 |
Mar 2002 |
US |
Continuation in Parts (1)
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Number |
Date |
Country |
Parent |
09069890 |
Apr 1998 |
US |
Child |
09276443 |
Mar 1999 |
US |