Polypeptide formulations and methods for making, using and characterizing them

• US 20060183178A1
• Filed: 02/17/2005
• Published: 08/17/2006
• Est. Priority Date: 02/17/2005
• Status: Active Grant

• Current Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Original Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Inventors: Chattaraj, Sarnath, Gulati, Poonam S., Levin, Eugene, Van Antwerp, William P., Minnoor, Elango S., Zhu, Xiao
• Granted Patent: US 7,241,586 B2
• Time in Patent Office: Days
• US Class Current: 435/14
• CPC Class Codes: A61B 2562/12 Manufacturing methods specially adapted for producing sensors for in-vivo measurements
• CPC Class Codes: A61B 5/1486 using enzyme electrodes, e.g. with immobilised oxidase
• CPC Class Codes: A61K 38/443 acting on CH-OH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 2333/904 acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/06 Preparation
• CPC Class Codes: G01N 30/74 Optical detectors measurement of intensity, velocity, spectral content, polarisation, or phase of infrared, visible or ultraviolet light G01J

Abstract

Embodiments of the invention include polypeptide formulations and methods for making, using and characterizing them. Embodiment of the invention include stabilized polypeptide formulations, for example stable glucose oxidase formulations that can be used with glucose sensors used in the management of diabetes. Another embodiment of the invention includes methods to characterize the concentration of nonionic surfactants in stabilized polypeptide formulation for example stable insulin formulations that can be used in the treatment of diabetes.

Claims

1. A method of determining the concentration of a non-ionic surfactant in an aqueous solution comprising the steps of:

(a) subjecting the aqueous solution to a chromatographic separation step; and

(b) analyzing the aqueous solution to from (b) via evaporative light scattering,

such that the concentration of the non-ionic surfactant in the aqueous solution is determined.

2. The method of claim 1, wherein the non-ionic surfactant is a poloxamer.

3. The method of claim 1, wherein the non-ionic surfactant is poloxamer 171, Triton X-100, Triton X-405, Triton BRIJ-35, Tween-20 or Tween-80.

4. The method of claim 1, wherein the non-ionic surfactant is extracted from the aqueous solution prior to the chromatographic separation step.

5. The method of claim 4, wherein the non-ionic surfactant extracted from the aqueous solution is concentrated prior to the chromatographic separation step.

6. The method of claim 1, wherein the chromatographic separation step comprises reverse-phase chromatography.

7. The method of claim 1, wherein the chromatographic separation step comprises high pressure liquid chromatography.

8. The method of claim 1, wherein the aqueous solution comprises a pharmaceutically acceptable composition.

9. The method of claim 1, wherein the aqueous solution comprises a therapeutic polypeptide.

10. The method of claim 1, wherein the aqueous solution comprises insulin.

11. The method of claim 1, wherein the concentration of the non-ionic surfactant in the aqueous solution is less than the critical micelle concentration.

12. The method of claim 1, wherein the concentration of the non-ionic surfactant in the aqueous solution is between about 0.1 and about 100 parts per million (ppm).

13. The method of claim 1, wherein the concentration of the non-ionic surfactant in the aqueous solution is between about 1 and about 20 parts per million (ppm).

14. A method of determining the concentration of a non-ionic surfactant in an aqueous solution comprising:

(a) extracting the non-ionic surfactant from the aqueous solution;

(b) concentrating the non-ionic surfactant from (a); and

(c) analyzing the concentrated non-ionic surfactant from (b), wherein the concentrated non-ionic surfactant is analyzed via high performance liquid chromatography coupled to an evaporative light scattering detector,

such that the concentration of the non-ionic surfactant in the aqueous solution is determined.

15. A composition comprising:

(a) glucose oxidase, wherein the glucose oxidase is present in a concentration of about 90 KU/mL to about 110 KU/mL;

(b) potassium sorbate, wherein the potassium sorbate is present in a concentration of about 0.12% w/v to about 0.15% w/v; and

(c) a potassium phosphate buffer;

wherein the potassium phosphate buffer is present in a concentration of about 0.01 M.

16. The composition of claim 15, wherein the glucose oxidase is stable for at least 6 months in a plastic container.

17. The composition of claim 15, wherein the glucose oxidase is present in a concentration of about 100 KU/mL.

18. The composition of claim 15, wherein the potassium sorbate is present in a concentration of about 0.15% w/v

19. A method of making a glucose oxidase composition that is stable for at least 6 months in a plastic container, the method comprising:

(a) preparing a glucose oxidase solution; and

(b) concentrating the glucose oxidase solution (a);

wherein the resulting concentrated glucose oxidase solution comprises glucose oxidase having a concentration of about 90 KU/mL to about 110 KU/mL, potassium sorbate having a concentration of 0.12% w/v to about 0.18% w/v; and

a potassium phosphate buffer having a concentration of about 0.01 M, so that the glucose oxidase composition that is stable for at least 6 months in a plastic container is made.

20. The method of claim 19, wherein the glucose oxidase solution is concentrated by a process comprising solid phase extraction or by a process comprising chromatography.

21. A method of making a sensor apparatus for implantation within a mammal comprising the steps of:

providing a base layer;

forming a conductive layer on the base layer, wherein the conductive layer includes a working electrode;

forming an analyte sensing layer on the conductive layer using a composition comprising glucose oxidase having a concentration of about 90 KU/mL to about 110 KU/mL, potassium sorbate having a concentration of about 0.12% w/v to about 0.18% w/v; and

a potassium phosphate buffer having a concentration of about 0.01 M;

optionally forming a protein layer on the analyte sensing layer;

forming an adhesion promoting layer on the analyte sensing layer or the optional protein layer;

forming an analyte modulating layer disposed on the adhesion promoting layer, wherein the analyte modulating layer includes a composition that modulates the diffusion of the analyte therethrough; and

forming a cover layer disposed on at least a portion of the analyte modulating layer, wherein the cover layer further includes an aperture over at least a portion of the analyte modulating layer.

22. A sensor made by the method of claim 21.