Application of Fiber-Optic Attenuated Total Reflection-FT-IR Methods for In Situ Characterization of Protein Delivery Systems in Real Time

Abstract

A fiber-optic coupled attenuated total reflection (ATR)-FT-IR spectroscopy technique was applied to the study of two different therapeutic delivery systems, acid degradable hydrogels and nanoparticles. Real time exponential release of a model protein, human serum albumin (HSA), was observed from two different polymeric hydrogels formulated with a pH sensitive cross-linker. Spectroscopic examination of nanoparticles formulated with an acid degradable polymer shell and encapsulated HSA exhibited vibrational signatures characteristic of both particle and payload when exposed to lowered pH conditions, demonstrating the ability of this methodology to simultaneously measure phenomena arising from a system with a mixture of components. In addition, thorough characterization of these pH sensitive delivery vehicles without encapsulated protein was also accomplished in order to separate the effects of the payload during degradation. When in situ, real time detection in combination with the ability to specifically identify different components in a mixture without involved sample preparation and minimal sample disturbance is provided, the versatility and suitability of this type of experiment for research in the pharmaceutical field is demonstrated.