Multicoated or multilayer entrapment matrix for protein biosensor

• US 20040234962A1
• Filed: 05/02/2003
• Published: 11/25/2004
• Est. Priority Date: 05/02/2003
• Status: Abandoned Application

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Jacobson, Ross W., Alarcon, Javier, Sherman, Douglas B., Pitner, J. Bruce, Hsieh, Helen V., Rowley, Jon A.
• Application Number:
  US10/428,295
• Publication Number:
  US 20040234962A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: G01N 33/5436 with ligand physically entrapped within the solid phase liposomes G01N33/5432; immunological test elements G01N33/54386
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92

Abstract

Multicoated or multilayer matrices capable of embedding, encapsulating or entrapping binding proteins, optionally with a reporter group attached, specific for analytes of interest, in particular sugars, and methods of making and using such matrices.

Claims

1. A multicoated or multilayer matrix capable of entrapping, embedding or encapsulating a binding protein comprising:

a core capable of physically or chemically entrapping a binding protein, and

one or more of;

a containment layer that ensures the integrity of said core,

an outer layer selectively permeable to an analyte of interest, or

a layer that is biocompatible.

2. The matrix of claim 1, comprising a

core capable of physically or chemically entrapping a binding protein,

a containment layer that ensures the integrity of the core, and

an outer layer selectively permeable to an analyte of interest.

3. The matrix of claim 2, comprising the core and a layer that is biocompatible.

4. The matrix of claim 1, wherein said core comprises a cross-linkable polymer.

5. The matrix of claim 4 wherein said cross-linkable polymer is alginate or hyaluronate.

6. The matrix of claim 1, wherein said containment layer comprises a compound selected from the group consisting of poly-L-lysine, poly-D-lysine, low and high molecular weight polyvinyl alcohols, SbQ-PVA, and Nafion®

.

7. The matrix of claim 1 wherein said outer layer comprises a compound selected from the group consisting of SbQ-PVA, HEMA, PVA, alginate and inorganic or organically modified inorganic sol-gels.

8. The matrix of claim 2 wherein said fourth layer comprises a compound selected from the group consisting of Nafion®

, polyethylene glycol (PEG), poly (2-hydroxyethyl methacrylate) (pHEMA), and alginate

9. The matrix of claim 1, wherein said core comprises a binding protein.

10. The matrix of claim 9, wherein said binding protein is physically entrapped in said core.

11. The matrix of claim 9, wherein said binding protein is covalently bound to said core.

12. The matrix of claim 9 wherein said binding protein is a sugar binding protein.

13. The matrix of claim 12, wherein said sugar binding protein is a mutated glucose/galactose or maltose binding protein.

14. The matrix of claim 9, wherein said binding protein additionally comprises a reporter group.

15. The matrix of claim 14, wherein said reporter group is a luminescent or fluorescent group.

17. A method for measuring a concentration of an analyte in a solution comprising the steps of:

(a) contacting the matrix of claim 14 with a solution;

(b) exposing the matrix to an energy source capable of producing a detectable signal therefrom;

(c) measuring the detectable signal; and

(d) correlating the measure of the detectable signal with the concentration of analyste in the solution.

18. The method of claim 17, wherein said analyte is a sugar.

20. The method of claim 17, wherein the detectable signal is measured episodically, continuously, or programmed.

21. A biosensor comprising:

the multilayered or multicoated matrix of claim 1 with at least one binding protein specific for an analyte of interest embedded, entrapped or encapsulated therein; and

a transducing element.

16. A method of making a multicoated or multilayer matrix comprising the steps of:

(a) forming a core;

(b) contacting the core with a first coating solution for a sufficient time to coat or layer the core;

(c) isolating and drying the first coating on the core to form the containment layer or coating;

(d) contacting the containment layer or coating with a second coating solution to form a multi-coated three dimensional structure; and

(e) isolating and drying the second coating on the multi-coated three dimensional structure to obtain a multicoated or multilayered matrix.

19. The method of claim 16 or 17, wherein the core of said matrix comprises alginate or hyaluronate, the containment layer comprises poly-L-lysine, and the outer layer comprises inorganic or organically modified inorganic sol-gel.

22. A method of making a biosensor comprising the steps of:

(a) combining a binding protein with a reporter group attached thereto with an embedding, encapsulating or entrapping compound to form a core;

(b) contacting the core with a first coating solution for a sufficient time to coat or layer the core;

(c) isolating and drying the first coating solution to form a containment layer;

(d) contacting the containment layer with a second coating solution for a sufficient time to coat or layer the containment layer;

(e) isolating and drying the second coating solution to form a outer layer to obtain a biosensor.