التفاصيل البيبلوغرافية
| العنوان: |
FUNCTIONALIZED BIOCHIPS FOR SPR-MS COUPLING |
| Document Number: |
20100285512 |
| تاريخ النشر: |
November 11, 2010 |
| Appl. No: |
12/734854 |
| Application Filed: |
November 27, 2008 |
| مستخلص: |
The invention relates to a method for coupling in-line the analysis of molecular interactions by surface plasmon resonance (SPR) with a structural identification by mass spectrometry using the same functionalized support for both types of analysis. |
| Inventors: |
Daniel, Régis (Issy Les Moulineaux, FR); Gonnet, Florence (Villiers Sur Orge, FR); Buchmann, William (Sainte Genevieve Des Bois, FR); Bellon, Sophie (Paris, FR); Jarroux, Nathalie (Ballainvilliers, FR); Anger-Leroy, Marielle (Les Ulis, FR) |
| Claim: |
1. A method for functionalizing the metal face of a support for analysis by surface plasmon resonance, said method comprising grafting a self-assembled monolayer of poly(ethylene oxide) directly onto the metal face of said support, in which the metal of the metal face is gold, and in which the poly(ethylene oxide) is a compound of formula (I) A-(CH2)n—(O—CH2—CH2)x-D (I) in which: n is equal to 1 or 2; x is an integer between 5 and 16; A is a group for anchoring the PEO onto the gold surface of the support by a strong interaction; and D is an optionally modified group for the binding of biomolecules. |
| Claim: |
2. The method according to claim 1, in which n is equal to 2. |
| Claim: |
3. The method according to claim 1, in which x is equal to 8. |
| Claim: |
4. The method according to claim 1, in which A corresponds to an —SH group. |
| Claim: |
5. The method according to claim 1, in which the PEO is O-(2-mercaptoethyl)-O′-(2-carboxyethy)heptaethylene glycol of formula HS—CH2—CH2—(O—CH2—CH2)8—COOH. |
| Claim: |
6. The method according to claim 1, which method comprises the series of following steps: 1) prior cleaning of the support; 2) grafting of the PEO onto the support; and 3) optionally, modification of group D of the PEO. |
| Claim: |
7. The method according to claim 6, in which the cleaning is carried out by UV-ozone treatment. |
| Claim: |
8. The method according to claim 6, in which group D represents a —COOH group which is modified in step 3) so as to give an N-hydroxysuccinimide group. |
| Claim: |
9. The method according to claim 6, in which the grafting is carried out by immersing the support in a vessel containing the poly(ethylene oxide) to be grafted, in solution. |
| Claim: |
10. A functionalized support for analysis by surface plasmon resonance, comprising a metal face onto which a self-assembled monolayer of poly(ethylene oxide) is directly grafted, in which the metal face is made of gold, and in which the poly(ethylene oxide) is a compound of formula (I) A-(CH2)n—(O—CH2—CH2)x-D (I) in which: n is equal to 1 or 2; x is an integer between 5 and 16; A is a group for anchoring the PEO onto the gold surface of the support by a strong interaction; and D is an optionally modified group for the binding of biomolecules. |
| Claim: |
11. The functionalized support produced by the method according to claim 1. |
| Claim: |
12. The functionalized support according to claim 10, further comprising “receptor” molecules immobilized on said support via a bond by means of grafted poly(ethylene oxide). |
| Claim: |
13. A method of surface plasmon resonance comprising a functionalized support according to claim 10, in particular in an surface plasmon resonance imaging (SPRi) experiment. |
| Claim: |
14. The method according to claim 13, said method comprising linking “receptor” molecules to the surface of said support and studying the molecular interactions of these molecules by surface plasmon resonance, in particular by SPRi. |
| Claim: |
15. A method of mass spectrometry comprising a functionalized support according to claim 10, in order to structurally identify analytes which have been specifically retained by receptor molecules covalently linked to said functionalized support. |
| Claim: |
16. The method according to claim 15, in which the mass spectrometry is carried out with MALDI-type ionization. |
| Claim: |
17. The method of a functionalized support according to claim 11, for carrying out two consecutive analyses: an analysis by SPR; then an analysis by mass spectrometry. |
| Claim: |
18. The method according to claim 17, in which the analysis by surface plasmon resonance is an analysis by surface plasmon resonance imaging. |
| Claim: |
19. The method according to claim 17, in which the analysis by MS is an analysis of MALDI type, in particular an analysis of MALDI-TOF type. |
| Claim: |
20. A method for coupling an analysis by SPR, in particular SPRi, with an analysis by MALDI MS or MS/MS, comprising: 1) immobilization of one or more “receptor” molecules on a functionalized support according to claim 10; then 2) placing of the support in an SPR analyzer and analysis, by SPR, of the interactions between the “receptor” molecule(s) immobilized and a sample of analytes; then 3) removal of the support from the SPR analyzer and placing of said support in a mass spectrometer, and structural analysis, by MS, of the analytes specifically retained by the “receptor” molecules during the SPR analysis. |
| Claim: |
21. A method for coupling an analysis by SPR, in particular SPRi, with an analysis by MS, in particular MALDI MS, or MS/MS comprising: 1) immobilization of one or more “receptor” molecules on a functionalized support according to claim 10; then 2) placing of the support in an SPR analyzer and an analysis, by SPR, of the interactions between the “receptor” molecule(s) immobilized and a sample of analytes; 3) in situ localized enzyme digestion of the analytes retained on the spots of the “receptor” molecules immobilized on the functionalized support, then placing of the support in a mass spectrometer, and structural analysis, by MS, or MS/MS, of the products of digestion of the analyte(s) present on the functionalized support. |
| Claim: |
22. A method for coupling an analysis by SPR, in particular SPRi, with an analysis by MS, in particular MALDI MS, comprising: 1) immobilization of one or more “receptor” molecules on a functionalized support according to claim 10; then 2) placing of the support in an SPR analyzer and analysis, by SPR, of the interactions between the “receptor” molecule(s) immobilized and a sample of analytes; then 3) removal of the support from the SPR analyzer and placing of said support in a mass spectrometer, and structural analysis, by MS, of the analytes specifically retained by the “receptor” molecules during the SPR analysis; then 4) in situ localized enzyme digestion of the analytes retained on the spots of the “receptor” molecules immobilized on the functionalized support, and placing of the support in a mass spectrometer, and structural analysis, by MALDI MS or MALDI MS/MS, of the products of digestion of the molecule(s) present on the functionalized support. |
| Current U.S. Class: |
435/18 |
| Current International Class: |
12; 01; 12; 05; 05; 01; 01 |
| رقم الانضمام: |
edspap.20100285512 |
| قاعدة البيانات: |
USPTO Patent Applications |
