Determining Spectroscopic Quantitation Limits for Misfolded Structures

Journal of Pharmaceutical Sciences | September 12, 2019

Brent S. Kendrick, John P. Gabrielson, Caroline Warly Solsberg, Eugene Ma, Libo Wang

J Pharm Sci. 2020 Jan;109(1):933-936

DOI: https://doi.org/10.1016/j.xphs.2019.09.004

Abstract

Protein secondary structures are frequently assessed using infrared and circular dichroism spectroscopies during drug development (e.g., during product comparability and biosimilarity studies, reference standard characterization, etc.) However, there is little information on the lower limits of quantitation of structural misfolds and impurities for these methods. A model system using a monoclonal antibody reference material was spiked at various levels with a protein that had a significantly different secondary structure to represent the presence of a stable and discreet structural misfold. The ability of circular dichroism, transmission Fourier transform infrared spectroscopy and microfluidic modulation spectroscopy, along with various spectral comparison algorithms, were assessed for their ability to detect the presence and quantify the amount of the misfolded structure.