WO2007121436B1 - Methods, particles, and kits for determining activity of a kinase - Google Patents
Methods, particles, and kits for determining activity of a kinaseInfo
- Publication number
- WO2007121436B1 WO2007121436B1 PCT/US2007/066781 US2007066781W WO2007121436B1 WO 2007121436 B1 WO2007121436 B1 WO 2007121436B1 US 2007066781 W US2007066781 W US 2007066781W WO 2007121436 B1 WO2007121436 B1 WO 2007121436B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- fluorescent
- particle
- particles
- support substrate
- reporter
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
- C12Q1/485—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase involving kinase
Abstract
Methods, particles and kits for determining kinase activity within a sample are provided. An embodiment of a method includes exposing a fluorescent particle to an assay, wherein the fluorescent particle includes a support substrate having one or more fluorescent materials and a peptide substrate coupled to the support substrate via a functional group of the support substrate. The method further includes phosphorylating the peptide substrate during exposure of the fluorescent particle to the assay and processing the fluorescent particle such that the peptide substrate is dephosphorylated and a polarized double bond is generated at a dephosphorylated site. In addition, the method includes coupling a fluorescent reporter having a nucleophilic terminal group to the fluorescent particle via the polarized double bond.
Claims
AMENDED CLAIMS received by the International Bureau on 28 December 2007 (28.12.07) WHAT IS CLAIMED IS:
1. A method, comprising: exposing a ftuoroscent particle to an assay, wherein the fluorescent particle comprises: a support substrate comprising one or more fluorescent materials; and a peptide substrate coupled to the support substrate via a functional group of the support substrate; phosphorylaling the peptide substrate during the step of exposing the fluorescent particle to the assay; processing the fluorescent particle such that the peptide substrate is dephosphorylated and a polarized double bond is generated at a dephosphorylated site; and coupling a fluorescent reporter comprising a nucleophilic terminal group to the fluorescent particle via the polarized double bond.
2. The method of claim I, wherein the fluorescent reporter comprises: a fluorescent compound; and one or more spacer compounds interposed between the nuclcophihic terminal group and the fluorescent compound.
3. The method of claim 2, wherein the one or more spacer compounds collectively comprise between approximately 1 atom and approximately 25 atoms.
4. The method of claim 1, wherein the step of coupling the fluorescent reporter to the fluorescent particle comprises coupling a fluorescent reporter that is configured to emit fluorescence within a different wavelength range than the one or more fluorescent materials of the support substrate.
5. The method of claim 4, wherein the fluorescent reporter is configured to emit fluorescence at a wavelength greater than approximately 500 nm.
6. The method of claim 1, wherein the step of phosphorylating the peptide substrate comprises exposing the fluorescent particle to an ionic liquid heated via microwaves.
7. The method of claim 1 , wherein the step of processing the fluorescent particle comprises exposing the fluorescent particle to an ionic liquid.
8. The method of claim 7, wherein the step of processing the fluorescent particle further comprises microwave heating the fluorescent particle and the ionic liquid.
9. The method of claim 1 , wherein the step of coupling the fluorescent reporter comprises exposing the fluorescent particle to an ionic liquid.
10. The method of claim 9, wherein the step of coupling the fluorescent reporter further comprises microwave heating the fluorescent particle and the ionic liquid.
11. A particle, comprising: a support substrate comprising one or more fluorescent materials; and a kinase-specific peptide substrate coupled to the support substrate via a functional group of the support substrate.
12. The particle of claim 11, wherein the kinase-specific peptide substrate is a phosphorylated peptide substrate,
13. The particle of claim 11 , wherein the kinase-specific peptide substrate comprises a Michael acceptor.
14. The particle of claim 11, further comprising a fluorescent reporter coupled to the kinase- specific peptide substrate via a nucleophilic terminal group of the fluorescent reporter.
15. The particle of claim 14, wherein the fluorescent reporter comprises: a fluorescent compound; and one or more spacer compounds interposed between the nucleophilic terminal group and the fluorescent compound.
16. The particle of claim 15, wherein the one or more spacer compounds collectively comprise between approximately 1 atom and approximately 25 atoms.
17. The particle of claim 15, wherein the one or more spacer compounds collectively comprise between approximately 5 atoms and approximately 25 atoms.
18. The particle of claim 15, wherein the fluorescent compound is configured to emit fluorescence within a different wavelength range than the one or more fluorescent materials of the support substrate.
19. The particle of claim 18, wherein the fluorescent compound is configured to emit fluorescence at a wavelength between approximately 400 nm and approximately 580 nm.
20. The particle of claim 18, wherein the fluorescent compound is configured to emit fluorescence at a wavelength greater than approximately 500 nm.
21. The particle of claim 15, wherein the fluorescent compound comprises a hydrophilic fluorescent compound.
22. The particle of claim 11, wherein the support substrate is a crosslinkcd support substrate.
23. A method, comprising: exposing a pooled population of different subsets of fluorescent particles to a sample, wherein at least some of the fluorescent particles comprise: a support substrate comprising one or more fluorescent materials configured to emit fluorescence in a first wavelength range, and wherein at least some of the different subsets of fluorescent particles respectively comprise a different configuration of the one or more fluorescent materials; and a peptide substrate coupled to the support substrate via a functional gioup of the support substrate, and wherein at least some of the different subsets of fluorescent particles respectively comprise a different peptide substrate; exposing (he sample and the pooled population to a phosphorylation process configured to add phosphate groups to accepting residues of the peptide substrates; processing a plurality of the fluorescent particles subsequent to exposing the sample and the pooled population to the phosphorylation process such that if any phosphorylated peptide substrates exist among the plurality of fluorescent
particles, the phosphorylated peptide substrates are dephosphorylated and polarized double bonds are generated at dcphosphorylated sites of the peptide substrates; further processing the plurality of the fluorescent particles such that if any polarized double bonds exist among the dephosphorylated sites of the peptide substrates, fluorescent reporters are coupled to the fluorescent particles al positions of the polarized double bonds via nucleophilic terminal groups of the fluorescent reporters, wherein the fluorescent reporters arc configured to emit fluorescence in a second wavelength range distinct from the first wavelength range; measuring fluorescence emissions of the plurality of the fluorescent particles subsequent to further processing the plurality of the fluorescent particles; identifying subset classifications of the particles in the sample based upon measured fluorescence emissions within the first wavelength range; and determining, based upon the existence of or lack of measured fluorescence emissions within the second wavelength range, an amount of kinase activity within the sample when the sample and the pooled population are exposed to the phosphorylation process,
24. The method of claim 23, wherein the step of identifying subset classifications of the particles comprises determining the identity of more than approximately 100 subset classifications,
25. The method of claim 23, wherein the step of further processing the plurality of the fluorescent particles comprises: coupling a first fluorescent reporter to fluorescent particles within a first subset of the plurality of fluorescent particles; and coupling a different fluorescent reporter to fluorescent particles within a second subset of the plurality of fluorescent particles, and wherein the different fluorescent reporter is configured to emit fluorescence in a wavelength range distinct from a wavelength range the first fluorescent reporter is configured to emit.
26, A kit, comprising: a plurality of fluorescent particles, wherein each of the plurality of fluorescent particles comprises a support substrate with one or more fluorescent materials configured to emit flυorescence in a first wavelength range; and one or more kinase-specific peptide substrates coupled to at least some of the plurality of fluorescent particles via functional groups of the support substrates.
28, The kit of claim 26, wherein the one or more kinase-specific peptide substrates are respectively coupled to different subsets of the plurality of fluorescent particles.
31. Tho kit of Claim 26, further comprising; a phosphorylation reagent configured to phosphorylate the one or more kinase-specific peptide substrates; a beta-elimination reagent configured to dcphosphorylate the one or more kinase-specific peptide substrates and generate Michael acceptors at the dephosphorylation sites of the one or more kinase-specific peptide substrates; and one or more fluorescent reporter reagents each having a nucleophilic terminal group and one or more fluorescent compounds configured to emit fluorescence in a second wavelength range distinct from the first wavelength range.
32. The kit of claim 31, wherein at least one of the phosphorylation reagent, the beta- elimination reagent, and tho one or more fluorescent reporter reagents comprises an ionic liquid,
33. The kit of claim 31 , wherein at least one of the one or more fluorescent reporter reagents comprises a hydrophilic fluorescent compound.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP07782040A EP2013358A2 (en) | 2006-04-17 | 2007-04-17 | Methods, particles, and kits for determining activity of a kinase |
| JP2009506719A JP2009533073A (en) | 2006-04-17 | 2007-04-17 | Methods, particles and kits for determining kinase activity |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US74494906P | 2006-04-17 | 2006-04-17 | |
| US60/744,949 | 2006-04-17 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| WO2007121436A2 WO2007121436A2 (en) | 2007-10-25 |
| WO2007121436A3 WO2007121436A3 (en) | 2007-12-21 |
| WO2007121436B1 true WO2007121436B1 (en) | 2008-02-07 |
Family
ID=38541967
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US2007/066781 WO2007121436A2 (en) | 2006-04-17 | 2007-04-17 | Methods, particles, and kits for determining activity of a kinase |
Country Status (5)
| Country | Link |
|---|---|
| US (2) | US7795040B2 (en) |
| EP (1) | EP2013358A2 (en) |
| JP (1) | JP2009533073A (en) |
| CN (1) | CN101421414A (en) |
| WO (1) | WO2007121436A2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| HUE028235T2 (en) * | 2008-04-29 | 2016-12-28 | Psychemedics Corp | Solid phase multi-analyte assay |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5736330A (en) * | 1995-10-11 | 1998-04-07 | Luminex Corporation | Method and compositions for flow cytometric determination of DNA sequences |
| EP0852004B1 (en) * | 1995-10-11 | 2011-01-19 | Luminex Corporation | Multiplexed analysis of clinical specimens |
| US5981180A (en) * | 1995-10-11 | 1999-11-09 | Luminex Corporation | Multiplexed analysis of clinical specimens apparatus and methods |
| US6449562B1 (en) * | 1996-10-10 | 2002-09-10 | Luminex Corporation | Multiplexed analysis of clinical specimens apparatus and method |
| WO1998059233A1 (en) * | 1997-06-23 | 1998-12-30 | Luminex Corporation | Interlaced lasers for multiple fluorescence measurement |
| AU1080999A (en) * | 1997-10-14 | 1999-05-03 | Luminex Corporation | Precision fluorescently dyed particles and methods of making and using same |
| KR100593712B1 (en) * | 1998-01-22 | 2006-06-30 | 루미넥스 코포레이션 | Microparticles with Multiple Fluorescent Signals |
| US6046807A (en) * | 1998-05-14 | 2000-04-04 | Luminex Corporation | Diode laser based measurement apparatus |
| JP3946444B2 (en) * | 1998-05-14 | 2007-07-18 | ルミネックス コーポレイション | Configuration and method for zero dead time of flow cytometer |
| WO2001013119A1 (en) * | 1999-08-17 | 2001-02-22 | Luminex Corporation | Encapsulation of fluorescent particles |
| US7041453B2 (en) | 2002-08-22 | 2006-05-09 | Bioarray Solutions Ltd. | Molecular constructs and methods of use for detection of biochemical reactions |
| CN101072992A (en) * | 2004-11-12 | 2007-11-14 | 卢米尼克斯股份有限公司 | Methods and systems for positioning microspheres for imaging |
-
2007
- 2007-04-17 JP JP2009506719A patent/JP2009533073A/en not_active Withdrawn
- 2007-04-17 EP EP07782040A patent/EP2013358A2/en not_active Withdrawn
- 2007-04-17 US US11/736,254 patent/US7795040B2/en not_active Expired - Fee Related
- 2007-04-17 WO PCT/US2007/066781 patent/WO2007121436A2/en active Application Filing
- 2007-04-17 CN CNA2007800137002A patent/CN101421414A/en active Pending
-
2010
- 2010-08-23 US US12/861,521 patent/US20100317544A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| JP2009533073A (en) | 2009-09-17 |
| WO2007121436A2 (en) | 2007-10-25 |
| US7795040B2 (en) | 2010-09-14 |
| US20100317544A1 (en) | 2010-12-16 |
| US20080026403A1 (en) | 2008-01-31 |
| EP2013358A2 (en) | 2009-01-14 |
| WO2007121436A3 (en) | 2007-12-21 |
| CN101421414A (en) | 2009-04-29 |
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