METHOD FOR PRODUCING MOLECULE IMMOBILIZING SUBSTRATE, AND MOLECULE IMMOBILIZING SUBSTRATE

• US 20100233827A1
• Filed: 02/19/2010
• Published: 09/16/2010
• Est. Priority Date: 03/11/2009
• Status: Active Grant

• Current Assignee: Shin-Etsu Chemical Company Limited
• Original Assignee: Shin-Etsu Chemical Company Limited
• Inventors: ISHIHARA, Toshinobu, KUSAKI, Wataru, KINSHO, Takeshi, WATANABE, Takeru
• Granted Patent: US 9,034,661 B2
• Time in Patent Office: Days
• US Class Current: 436/528
• CPC Class Codes: C08L 83/04 Polysiloxanes
• CPC Class Codes: C08L 83/06 containing silicon bound to oxygen-containing groups C08L83/12 takes precedence
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Abstract

There is disclosed a method for producing a molecule immobilizing substrate, comprising at least the steps of:

• • forming, on a substrate, a monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film; and
  • chemically modifying the hydroxyl groups, cyano groups, or oxiranyl groups of the monomolecular film to transform them into carboxyl groups, to thereby form, on the substrate, the monomolecular film including the carboxyl groups, which are oriented toward an outmost surface of the monomolecular film.

There can be provided: a method for producing a molecule immobilizing substrate which is free of occurrence of an immobilized-molecule peeling problem in the case of conducting an assay by immobilizing molecules on the substrate.

Claims

1. A method for producing a molecule immobilizing substrate, comprising at least the steps of:

forming, on a substrate, a monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film; and

chemically modifying the hydroxyl groups, cyano groups, or oxiranyl groups of the monomolecular film to transform them into carboxyl groups, to thereby form, on the substrate, the monomolecular film including the carboxyl groups, which are oriented toward an outmost surface of the monomolecular film.

2. The method for producing a molecule immobilizing substrate according to claim 1, wherein the step of chemically modifying the hydroxyl groups to transform them into the carboxyl groups, is conducted by reacting a dicarboxylic anhydride or oxidizing agent with each of the hydroxyl groups.

3. The method for producing a molecule immobilizing substrate according to claim 1, wherein the step of chemically modifying the cyano groups to transform them into the carboxyl groups, is conducted by hydrolyzing the cyano groups.

4. The method for producing a molecule immobilizing substrate according to claim 1, wherein the step of chemically modifying the oxiranyl groups to transform them into the carboxyl groups, is conducted by reacting a thiol having a carboxyl group with each of the oxiranyl groups.

5. The method for producing a molecule immobilizing substrate according to claim 4, further comprising the step of conducting addition of a sulfonic acid, in the step of reacting the thiol having a carboxyl group with each of the oxiranyl groups.

6. The method for producing a molecule immobilizing substrate according to claim 1, further comprising the step of reacting an acid with hydroxyl-group precursory functional groups to form, on the substrate, the monomolecular film including the hydroxyl groups, which are oriented toward an outmost surface of the monomolecular film, in the step of forming the monomolecular film on the substrate.

7. The method for producing a molecule immobilizing substrate according to claim 2, further comprising the step of reacting an acid with hydroxyl-group precursory functional groups to form, on the substrate, the monomolecular film including the hydroxyl groups, which are oriented toward an outmost surface of the monomolecular film, in the step of forming the monomolecular film on the substrate.

8. The method for producing a molecule immobilizing substrate according to claim 6, wherein the hydroxyl-group precursory functional groups are each:

an alkoxymethoxy group including an alkoxyl group moiety having 1 to 6 carbon atoms; and

/or an oxiranyl group.

9. The method for producing a molecule immobilizing substrate according to claim 7, wherein the hydroxyl-group precursory functional groups are each:

an alkoxymethoxy group including an alkoxyl group moiety having 1 to 6 carbon atoms; and

/or an oxiranyl group.

10. The method for producing a molecule immobilizing substrate according to claim 1, wherein the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, is attained by immersing the substrate in a solution containing a silane compound having a hydroxyl-group precursory functional group, cyano group, or oxiranyl group.

11. The method for producing a molecule immobilizing substrate according to claim 2, wherein the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, is attained by immersing the substrate in a solution containing a silane compound having a hydroxyl-group precursory functional group, cyano group, or oxiranyl group.

12. The method for producing a molecule immobilizing substrate according to claim 3, wherein the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, is attained by immersing the substrate in a solution containing a silane compound having a hydroxyl-group precursory functional group, cyano group, or oxiranyl group.

13. The method for producing a molecule immobilizing substrate according to claim 4, wherein the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, is attained by immersing the substrate in a solution containing a silane compound having a hydroxyl-group precursory functional group, cyano group, or oxiranyl group.

14. The method for producing a molecule immobilizing substrate according to claim 10, wherein the silane compound having a hydroxyl-group precursory functional group, cyano group, or oxiranyl group is a silane compound represented by the following general formula (1).

Y₃Si—

(CH₂)_m—

Z 

 

(1)

(In the formula, “

m”

represents an integer of 3 to 16;

“

Z”

represents a hydroxyl-group precursory functional group, cyano group, or oxiranyl group; and

each “

Y”

independently represents a halogen atom, or an alkoxyl group having 1 to 4 carbon atoms).

15. The method for producing a molecule immobilizing substrate according to claim 11, wherein the silane compound having a hydroxyl-group precursory functional group, cyano group, or oxiranyl group is a silane compound represented by the following general formula (1).

Y₃Si—

(CH₂)_m—

Z 

 

(1)

(In the formula, “

m”

represents an integer of 3 to 16;

“

Z”

represents a hydroxyl-group precursory functional group, cyano group, or oxiranyl group; and

each “

Y”

independently represents a halogen atom, or an alkoxyl group having 1 to 4 carbon atoms).

16. The method for producing a molecule immobilizing substrate according to claim 12, wherein the silane compound having a hydroxyl-group precursory functional group, cyano group, or oxiranyl group is a silane compound represented by the following general formula (1).

Y₃Si—

(CH₂)_m—

Z 

 

(1)

(In the formula, “

m”

represents an integer of 3 to 16;

“

Z”

represents a hydroxyl-group precursory functional group, cyano group, or oxiranyl group; and

each “

Y”

independently represents a halogen atom, or an alkoxyl group having 1 to 4 carbon atoms).

17. The method for producing a molecule immobilizing substrate according to claim 13, wherein the silane compound having a hydroxyl-group precursory functional group, cyano group, or oxiranyl group is a silane compound represented by the following general formula (1).

Y₃Si—

(CH₂)_m—

Z 

 

(1)

(In the formula, “

m”

represents an integer of 3 to 16;

“

Z”

represents a hydroxyl-group precursory functional group, cyano group, or oxiranyl group; and

each “

Y”

independently represents a halogen atom, or an alkoxyl group having 1 to 4 carbon atoms).

18. The method for producing a molecule immobilizing substrate according to claim 14, wherein, in the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, by using the silane compound represented by the general formula (1) having the hydroxyl-group precursory functional group, cyano group, or oxiranyl group;

at least one of silane compounds represented by the following general formulae (2) and (3) is mixed with the silane compound represented by the general formula (1), and the monomolecular film is formed by using the mixture.

Y′

₃Si—

(CH₂)_n—

CH₃ 

 

(2)

Y′

₃Si—

(CH₂)_n—

OCH₃ 

 

(3)

(In the formula, “

n”

represents an integer of 0 to

“

m”

represents the value in the general formula (1); and

each “

Y′

”

independently represents a halogen atom, or an alkoxyl group having 1 to 4 carbon atoms).

19. The method for producing a molecule immobilizing substrate according to claim 15, wherein, in the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, by using the silane compound represented by the general formula (1) having the hydroxyl-group precursory functional group, cyano group, or oxiranyl group;

at least one of silane compounds represented by the following general formulae (2) and (3) is mixed with the silane compound represented by the general formula (1), and the monomolecular film is formed by using the mixture.

Yi₃Si—

(CH₂)_n—

CH₃ 

 

(2)

Y′

₃Si—

(CH₂)_n—

OCH₃ 

 

(3)

(In the formula, “

n”

represents an integer of 0 to “

m”

;

“

m”

represents the value in the general formula (1); and

each “

Y′

”

independently represents a halogen atom, or an alkoxyl group having 1 to 4 carbon atoms).

20. The method for producing a molecule immobilizing substrate according to claim 16, wherein, in the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, by using the silane compound represented by the general formula (1) having the hydroxyl-group precursory functional group, cyano group, or oxiranyl group;

at least one of silane compounds represented by the following general formulae (2) and (3) is mixed with the silane compound represented by the general formula (1), and the monomolecular film is formed by using the mixture.

Yi₃Si—

(CH₂)_n—

CH₃ 

 

(2)

Y′

₃Si—

(CH₂)_n—

OCH₃ 

 

(3)

(In the formula, “

n”

represents an integer of 0 to W_m;

“

m”

represents the value in the general formula (1); and

each “

Y′

”

independently represents a halogen atom, or an alkoxyl group having 1 to 4 carbon atoms).

21. The method for producing a molecule immobilizing substrate according to claim 17, wherein, in the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, by using the silane compound represented by the general formula (1) having the hydroxyl-group precursory functional group, cyano group, or oxiranyl group;

at least one of silane compounds represented by the following general formulae (2) and (3) is mixed with the silane compound represented by the general formula (1), and the monomolecular film is formed by using the mixture.

Y′

₃Si—

(CH₂)_n—

CH₃ 

 

(2)

Y′

₃Si—

(CH₂)_n—

OCH₃ 

 

(3)

(In the formula, “

n”

represents an integer of 0 to “

m”

;

“

m”

represents the value in the general formula (1); and

each “

Y′

”

independently represents a halogen atom, or an alkoxyl group having 1 to 4 carbon atoms).

22. The method for producing a molecule immobilizing substrate according to claim 14, wherein, in the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, by using the silane compound represented by the general formula (1) having the hydroxyl-group precursory functional group, cyano group, or oxiranyl group;

a catalyst to be mixed with the silane compound represented by the general formula (1) comprises an organic base.

23. The method for producing a molecule immobilizing substrate according to claim 18, wherein, in the step of forming, on the substrate, the monomolecular film including hydroxyl groups, cyano groups, or oxiranyl groups, which are oriented toward an outmost surface of the monomolecular film, by using the silane compound represented by the general formula (1) having the hydroxyl-group precursory functional group, cyano group, or oxiranyl group;

a catalyst to be mixed with the silane compound represented by the general formula (1) comprises an organic base.

24. The method for producing a molecule immobilizing substrate according to claim 22, wherein the organic base is a pyrrolidine derivative or piperidine derivative.

25. The method for producing a molecule immobilizing substrate according to claim 23, wherein the organic base is a pyrrolidine derivative or piperidine derivative.

26. The method for producing a molecule immobilizing substrate according to claim 1, wherein the molecule immobilizing substrate is used to immobilize a biomolecule thereon.

27. The method for producing a molecule immobilizing substrate according to claim 25, wherein the molecule immobilizing substrate is used to immobilize a biomolecule thereon.

28. The method for producing a molecule immobilizing substrate according to claim 26, wherein the biomolecule is a nucleic acid or protein.

29. The method for producing a molecule immobilizing substrate according to claim 27, wherein the biomolecule is a nucleic acid or protein.

30. A molecule immobilizing substrate produced by the method for producing a molecule immobilizing substrate according to claim 1.

31. A molecule immobilizing substrate produced by the method for producing a molecule immobilizing substrate according to claim 29.