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1. (WO2017001374) IMMOBILISATION EN SURFACE D'UNE MOLÉCULE DE RECONNAISSANCE D'ANALYTE
Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

A method for immobilizing an analyte-recognizing molecule (1 ) on a surface (2') functionalized with chemical groups Y1 suitable for reacting with a chemical group X2 of a coupling molecule (7) to form a reaction product comprising a chemical group Y2 suitable for reacting with the analyte-recognizing molecule (1 ), the method comprising the steps of: a. Providing the functionalized surface (2'),

b. Contacting the functionalized surface (2') with a solution (6) comprising simultaneously:

i. The coupling molecule (7), and

ii. The analyte-recognizing molecule (1 ).

The method according to claim 1 , wherein the analyte-recognizing molecule (1 ) comprises at least one group selected from -NH2 , -SH, -COOH, -OH, aldehyde, ketone, phosphate and pyrophosphate groups and wherein the coupling molecule (7) is suitable for reacting with the at least one group.

The method according to claim 2

- wherein the analyte-recognizing molecule (1 ) has a -NH2 group and Y2 is selected from the group consisting of o-acylisourea, N-hydroxysuccinimide ester, isothiocyanate, isocyanate, acyl azide, sulfonylchoride, aldehyde, oxirane, carboxyl, carbonate, arylating agents, imidoester, acid anhydride, fluorophenyl ester, and hydroxymethyl phosphine derivative, or

- wherein the analyte-recognizing molecule (1 ) has a -SH group and Y2 is selected from the group consisting of activated halogens, maleimide, aziridine, acryloyl, vinylsulfone, arylating agent and disulfide, or

- wherein the analyte-recognizing molecule (1 ) has a -COOH group and Y2 is selected from the group consisting of diazoalkane and diazoacetyl, or

- wherein the analyte-recognizing molecule (1 ) has a aldehyde or ketone group and Y2 is selected from the group consisting of hydrazide and -NH2, or

- wherein the analyte-recognizing molecule (1 ) has a phosphate or a pyrophosphate group and Y2 is carbonyldiimidazole.

The method according to claim 3, wherein the analyte-recognizing molecule (1 ) has a -NH2 group and Y2 is N-hydroxysuccinimide ester. The method according to any one of the preceding claims, wherein Y1 is selected from the group consisting of -N3, alkyne, and -COOH wherein

• If Y1 is -N3, X2 is an alkyne which is strained or not, and if the alkyne is not strained, step b is performed in presence of a catalyst suitable for enabling the reaction of an alkyne which is not strained

• if Y1 is an alkyne which is strained or not, X2 is -N3, and if the alkyne is not strained, step b is performed in presence of a catalyst suitable for enabling the reaction of an alkyne which is not strained

• if Y1 is -COOH, X2 is either a carbodiimide or a diazo group =N2. The method according to claim 5 wherein Y1 is -N3 and X2 is an alkyne group of a cycloalkyne.

The method according to claim 6 as depending on claim 4 wherein the coupling molecule (7) has the general formula:


8. The method according to any one of the preceding claims, wherein the solution (6) comprises the coupling molecule (7) at a concentration at least equal to 1 μΜ.

9. The method according to any one of the preceding claims, wherein the solution (6) comprises the coupling molecule (7) at a concentration of at most 1 mM, preferably at most 100 μΜ, more preferably at most 10 μΜ.

10. The method according to any one of the preceding claims, wherein the analyte-recognizing molecule (1 ) is present in the solution (6) at a concentration at least equal to 1 *10 2 mol/l.

1 1 . The method according to any one of the preceding claims, wherein the solution (6) comprising the analyte-recognizing molecule (1 ) and the coupling molecule (7) is prepared by mixing the analyte-recognizing molecule (1 ) and the coupling molecule (7) at most 2 hours and preferably at most 30 min before step b.

12. The method according to any one of the preceding claims, wherein the coupling molecule (7) has the general formula X2-R2- Y2 wherein R2 is an organic group, branched or not, saturated or not, comprising from 1 to 30 carbon atoms, optionally comprising one or more heteroatoms selected from the group consisting of -N-, -0-, and -S-, and optionally comprising one or more oxo substituents.

13. The method according to any one of the preceding claims, wherein step b comprises the deposition on the functionalized surface (2') of a drop of the solution (6).

14. The method according to any one of the preceding claims, wherein the functionalized surface (2') is coupled with a transducer for generating a readable signal output upon interaction of an analyte (10) with the immobilized analyte-recognizing molecule (1 ).

15. A solution (6) comprising simultaneously:

i. a coupling molecule (7) having a chemical group X2 suitable for reacting with a chemical group Y1 of a surface (2') to form a reaction product comprising a chemical group Y2 suitable for reacting with an analyte-recognizing molecule (1 ), and

ii. the analyte-recognizing molecule (1 ).

16. A kit of parts comprising:

a. a first solution (6') comprising a coupling molecule (7) having a chemical group X2 suitable for reacting with a chemical group Y1 of a surface (2') to form a reaction product comprising a chemical group Y2 suitable for reacting with an analyte-recognizing molecule (1 ), and,

b. a second solution (6") comprising the analyte-recognizing molecule (1 ),

wherein the first and the second solution are such that once mixed together they form a solution (6) comprising:

• the coupling molecule (7) at a concentration at least equal to 1 μΜ and at most equal to 1 mM,

• the analyte-recognizing molecule (1 ) at a concentration at least equal to 1 0"12 mol/l.

17. The kit of parts according to claim 15, further comprising a sensor for the detection of an analyte (10) in a sample fluid, the sensor comprising:

i. the surface (2') functionalized with chemical groups Y1 and suitable for immobilizing the analyte-recognizing molecule (1 ) by reacting the chemical groups Y1 with the coupling molecule (7) to form the reaction product suitable for reacting with the analyte-recognizing molecule (1 ), and ii. a transducer coupled with the functionalized surface (2') and suitable for generating a readable signal output upon interaction of the analyte (10) with the immobilized analyte-recognizing molecule (1 ).