Overcoming CD spectroscopy sample preparation challenges in protein structural analysis for biosimilars.

Agbelusi, Temitope; Pancholi, Ketan; Kumar, Harini Sampath; Kumar, Amit

Abstract

Circular dichroism (CD) spectroscopy is a valuable tool in the pharmaceutical industry for determining the secondary and tertiary structure, folding, stability, and interactions of proteins in biosimilars. However, protein aggregation due to its interaction with surfactants can introduce variations in CD spectra, leading to false interpretations. To investigate this effect, we prepared aqueous solutions of α-chymotrypsin with the same amount of surfactant but at different solution preparation steps. Interestingly, the CD spectra of these solutions showed slight differences despite having the same surfactant concentration. This suggests that the order of surfactant addition can influence protein conformation. To address this issue, we propose a method for preparing aqueous protein solutions that yields reproducible CD spectra. This method is expected to benefit the pharmaceutical industry by improving the accuracy of CD spectroscopy for protein structural analysis and evaluation of protein functionality.

Citation

AGBELUSI, T., PANCHOLI, K., KUMAR, H.S. and KUMAR, A. 2024. Overcoming CD spectroscopy sample preparation challenges in protein structural analysis for biosimilars. Preprints [online]. Available from: https://doi.org/10.20944/preprints202401.0441.v1

• Working Paper Type: Preprint
• Deposit Date: Feb 3, 2024
• Publicly Available Date: Feb 3, 2024
• DOI: https://doi.org/10.20944/preprints202401.0441.v1
• Keywords: Circular dichroism spectroscopy; Proteins; Protein structures; Pharmaceutics
• Public URL: https://rgu-repository.worktribe.com/output/2204936
• Additional Information: This file also includes supplementary materials that are posted separately from the main text on Preprints.