Analysis of the Structure of 14 Therapeutic Antibodies Using Circular Dichroism Spectroscopy

Overview

Journal Anal Chem

Specialty Chemistry

Date 2024 Sep 10

PMID 39255385

Authors

Maria G Bruque

Alison Rodger

Soren Vronning Hoffmann

Nykola C Jones

Jean Aucamp

Tim R Dafforn

Owen R T Thomas

Affiliations

Soon will be listed here.

Abstract

Understanding the impact of the manufacturing environment on therapeutic monoclonal antibody (mAb) structures requires new process analytical technology. Here, we describe the creation of a new reference set for the circular dichroism (CD) spectra of mAbs. Data sets of the highest quality were collected by synchrotron radiation CD for 14 different mAbs in both native and acid-stressed states. Deconvolution of far-UV spectra for the mAb cohort identified two current reference sets (SP175 and SMP180) as assigning accurate secondary structures, irrespective of the analysis program employed. Scrutiny of spectra revealed significant variation in the far-UV and especially near-UV CD of the 14 mAbs. Two spectral features were found to be sensitive to changes in solution pH, i.e., the far-UV positive peak at 201-202 nm and the near-UV negative exciton couplet around 230-240 nm. The latter feature offers attractive possibilities for in-line CD-based monitoring of the mAb structure during manufacture.

Citing Articles

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