Use of In-line Raman Spectroscopy as a Non-destructive and Rapid Analytical Technique to Monitor Aggregation of a Therapeutic Protein

摘要

Structural/chemical changes induced by thermal and shear stresses during manufacture and shipping as well as potential interactions with various contact surfaces may lead to protein aggregation. Liquid formulations are generally at a greater risk of physical degradation. There are well-established bio-analytical techniques to analyze protein aggregation, but these are often time-consuming, complex and may require special handling of the samples. Spectroscopic methods such as FTIR, Raman and NIR are becoming more popular as the instrumentation becomes more robust and user-friendly with the evolution of process analytical technology (PAT) in pharmaceutical industry. Although off-line Raman spectroscopic methods have been used to monitor structural/chemical changes of protein solutions [1,2], there are no published reports of the use of in-line Raman methods. Recently, aggregation of human insulin, a relatively moderate sized protein, was studied using an off-line Raman spectroscopy method [3], and the authors have also reported development of a multivariate calibration model for aggregation of human insulin under stress conditions. A similar approach is used in this study for a larger model globular protein.