Polypeptide formulations and methods for making, using and characterizing them

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.

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.