Polycationic quaternary ammonium polymer coatings for immobilizing biological samples

• US 8,617,895 B2
• Filed: 09/22/2005
• Issued: 12/31/2013
• Est. Priority Date: 09/23/2004
• Status: Active Grant

• Current Assignee: Tripath Imaging Incorporated (Becton, Dickinson & Co)
• Original Assignee: Tripath Imaging Incorporated (Becton, Dickinson & Co)
• Inventors: Fox, William Alan, Ray, William Carl III
• Primary Examiner(s): Haq, Shafiqul
• Assistant Examiner(s): Hollinden, Gary E
• Application Number:
  US11/233,496
• Publication Number:
  US 20060073510A1
• Time in Patent Office: 3,022 Days
• Field of Search: None
• US Class Current: 436/8
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• 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
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/108331 Preservative, buffer, anticoagulant or diluent
• CPC Class Codes: Y10T 436/173845 Amine and quaternary ammonium

Abstract

The present invention is directed to a pre-coated substrate, such as a slide, that is useful for immobilizing a sample. The invention is further provides methods of preparing such pre-coated substrates and methods of analyzing biological samples immobilized on such pre-coated substrate. The substrate is coated with a polycationic polymeric coating material specifically selected such that that coated substrate exhibits increased stability and prolonged shelf-life. Preferred polymeric coating materials include allylic or vinylic polymers having cationic groups thereon and having no more than a small percentage of peptidic monomeric linkages, particularly polydiallyldimethylammonium (PDDA).

Claims

1. A method for preparing a coated substrate adapted for immobilizing a biological sample, said method comprising:

providing a glass substrate having a surface comprising a plurality of anionic groups;

contacting the substrate with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric material comprising at least one monomer selected from the group consisting of diallyldimethylammonium, methylacrylamidopropyltrimethylammonium, methacryloyloxyethyltrimethylammonium, 4-vinyl-benzyltrimethylammonium, acryloxyethyldimethylbenzyl ammonium, acryloxyethyltrimethyl ammonium, methacryloxyethyldimethylammonium, methacryloxyethyltrimethylbenzylammonium, trimethyl-2-methacryloylethylammonium, trimethyl-2-methacrylaminopropylammonium, and mixtures thereof, said quaternary ammonium polymeric material present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of a single layer of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the substrate with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric coating material and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material;

wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da, and

drying the coating of the non-peptidic, quaternary ammonium polymeric material.

2. The method according to claim 1, wherein said step of drying the coated substrate is carried out at ambient temperature.

3. The method according to claim 2, wherein said step of drying the coated substrate is carried out for a period of time of about 5 minutes to about 1 hour.

4. The method according to claim 1, wherein said step of drying the coated substrate is carried out at a temperature of about 35°

C. to about 120°

C.

5. The method according to claim 4, wherein said step of drying the coated substrate is carried out for a period of time of about 1 minute to about 20 minutes.

6. The method according to claim 1, wherein the substrate is selected from the group consisting of slides, plates, beads, test tubes, cuvettes, and dipsticks.

7. The method according to claim 1, wherein the substrate is a microscope slide.

8. The method according to claim 1, wherein the non-peptidic, quaternary ammonium polymeric material comprises polydiallyldimethylammonium.

9. The method according to claim 1, wherein the non-peptidic, quaternary ammonium polymeric material comprises a copolymer of diallyldimethylammonium and at least one additional monomer.

10. The method of claim 1, wherein the non-peptidic, quaternary ammonium polymeric coating is present in solution at a concentration of about 0.15% (w/v) to about 0.75% (w/v).

11. The method according to claim 1, wherein the substrate surface has not been subjected to a cleaning process prior to contacting the substrate with the solution of the non-peptidic, quaternary ammonium polymeric material.

12. The method according to claim 11, wherein the substrate surface has not been subjected to a cleaning process incorporating an acid, a base, or an organic solvent prior to contacting the substrate with the solution of the non-peptidic, quaternary ammonium polymeric material.

13. The method according to claim 1, wherein the substrate surface has not be subjected to a silanization process prior to contacting the substrate with the solution of the non-peptidic, quaternary ammonium polymeric material.

14. The method according to claim 1, further comprising rinsing the substrate having the dried coating of the non-peptidic, quaternary ammonium polymeric material thereon.

34. The method of claim 1, wherein said concentration is about 0.25% (w/v).

15. A method for preparing a microscope slide adapted for immobilizing a biological sample, said method comprising:

providing a previously manufactured microscope slide having a surface comprising a plurality of anionic groups;

contacting the surface of the microscope slide with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric material present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the microscope slide with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric coating material and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material;

wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da,

drying the coating of the non-peptidic, quaternary ammonium polymeric material; and

rinsing the microscope slide having the dried coating of the non-peptidic, quaternary ammonium polymeric material thereon;

wherein the microscope slide surface has not been subjected to a cleaning process after receipt from the manufacturer and prior to contacting the microscope slide surface with the solution of the non-peptidic, quaternary ammonium polymeric material.

16. The method according to claim 15, wherein the slide comprises a material selected from the group consisting of glass, ceramics, and polymer materials.

17. The method according to claim 16, wherein the slide comprises a polymer selected from the group consisting of polystyrene, polyhydroxy methacrylate, polyethylene terephthalate, polytetrafluoroethylene, fluorinated ethylene, polydimethylsiloxane, copolymers or terpolymers thereof, and combinations thereof.

18. The method according to claim 15, wherein the non-peptidic, quaternary ammonium polymeric material comprises at least one monomer selected from the group consisting of diallyldimethylammonium, methylacrylamidopropyltrimethylammonium, methacryloyloxyethyltrimethylammonium, 4-vinyl-benzyltrimethylammonium, acryloxyethyldimethylbenzyl ammonium, acryloxyethyltrimethyl ammonium, methacryloxyethyldimethylammonium, methacryloxyethyltrimethylbenzylammonium, trimethyl-2-methacryloylethylammonium, trimethyl-2-methacrylaminopropylammonium, and mixtures thereof.

19. The method according to claim 15, wherein the non-peptidic, quaternary ammonium polymeric material comprises a polymer selected from the group of allylic polymers.

20. The method according to claim 15, wherein the non-peptidic, quaternary ammonium polymeric material comprises polydiallyldimethylammonium.

35. The method of claim 15, wherein said concentration is about 0.25% (w/v).

36. The method of claim 15, wherein said concentration is about 0.15% (w/v) to about 0.75% (w/v).

21. A method for preparing a microscope slide adapted for immobilizing a biological sample, said method comprising:

providing a previously manufactured microscope slide having a surface comprising a plurality of anionic groups; and

contacting the surface of the microscope slide with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the microscope slide with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric coating material and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material, wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da.

22. The method according to claim 21, wherein the slide comprises a material selected from the group consisting of glass, ceramics, and polymer materials.

23. The method according to claim 22, wherein the slide comprises a polymer selected from the group consisting of polystyrene, polyhydroxy methacrylate, polyethylene terephthalate, polytetrafluoroethylene, fluorinated ethylene, polydimethylsiloxane, copolymers or terpolymers thereof, and combinations thereof.

24. The method according to claim 21, wherein the non-peptidic, quaternary ammonium polymeric material comprises at least one monomer selected from the group consisting of diallyldimethylammonium, methylacrylamidopropyltrimethylammonium, methacryloyloxyethyltrimethylammonium, 4-vinyl-benzyltrimethylammonium, acryloxyethyldimethylbenzyl ammonium, acryloxyethyltrimethyl ammonium, methacryloxyethyldimethylammonium, methacryloxyethyltrimethylbenzylammonium, trimethyl-2-methacryloylethylammonium, trimethyl-2-methacrylaminopropylammonium, and mixtures thereof.

25. The method according to claim 21, wherein the non-peptidic, quaternary ammonium polymeric material comprises a polymer selected from the group of allylic polymers.

26. The method according to claim 21, wherein the non-peptidic, quaternary ammonium polymeric material comprises polydiallyldimethylammonium.

37. The method of claim 21, wherein said concentration is about 0.25% (w/v).

38. The method of claim 21, wherein said concentration is about 0.15% (w/v) to about 0.75% (w/v).

27. A method for preparing a microscope slide adapted for immobilizing a biological sample, said method comprising:

providing a previously manufactured microscope slide having a surface comprising a plurality of anionic groups; and

contacting the surface of the microscope slide with a composition comprising a solution of polydiallyldimethylammonium present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of the polydiallyldimethylammonium on at least a portion of the surface of the microscope slide with no intervening layers of a different coating material sandwiched between two or more layers of the polydiallyldimethylammonium and with no further layers of any material coated over said layer of the polydiallyldimethylammonium, wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da.

28. The method according to claim 27, wherein the slide comprises a material selected from the group consisting of glass, ceramics, and polymer materials.

29. The method according to claim 28, wherein the slide comprises a polymer selected from the group consisting of polystyrene, polyhydroxy methacrylate, polyethylene terephthalate, polytetrafluoroethylene, fluorinated ethylene, polydimethylsiloxane, copolymers or terpolymers thereof, and combinations thereof.

39. The method of claim 27, wherein said concentration is about 0.25% (w/v).

40. The method of claim 27, wherein said concentration is about 0.15% (w/v) to about 0.75% (w/v).

30. A method for preparing a coated substrate adapted for immobilizing a biological sample, said method comprising:

providing a glass substrate having a surface comprising a plurality of anionic groups; and

contacting the surface of the glass substrate with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric material present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the glass substrate microscope slide with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric material, and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material, wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da.

31. The method according to claim 30, wherein the substrate is selected from the group consisting of slides, plates, beads, test tubes, cuvettes, and dipsticks.

32. The method of claim 30, wherein said concentration is about 0.25% (w/v).

33. The method of claim 30, wherein said concentration is about 0.15% (w/v) to about 0.75% (w/v).

41. A method for preparing a coated substrate adapted for immobilizing a biological sample, said method comprising:

providing a glass substrate having a surface comprising a plurality of anionic groups;

contacting the substrate with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric material comprising at least one monomer selected from the group consisting of diallyldimethylammonium, methylacrylamidopropyltrimethylammonium, methacryloyloxyethyltrimethylammonium, 4-vinyl-benzyltrimethylammonium, acryloxyethyldimethylbenzyl ammonium, acryloxyethyltrimethyl ammonium, methacryloxyethyldimethylammonium, methacryloxyethyltrimethylbenzylammonium, trimethyl-2-methacryloylethylammonium, trimethyl-2-methacrylaminopropylammonium, and mixtures thereof, said quaternary ammonium polymeric material present in a concentration of about 0.15% (w/v) to about 0.75% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of a single layer of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the substrate with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric coating material and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material and

drying the coating of the non-peptidic, quaternary ammonium polymeric material.