Application of NMR and Chemometric Analyses to Better Understand the Quality Attributes in pH and Thermally Degraded Monoclonal Antibodies.

Autor: Beaumont VA; Pfizer, Inc. BioTherapeutics Pharmaceutical Sciences, Analytical Research and Development, 1 Burtt Road, Andover, MA, 01810, USA. victor.beaumont@pfizer.com.; Pfizer, Inc. Pharmaceutical Sciences Small Molecules, Analytical Research and Development, Discovery Park, Ramsgate Road, Sandwich, CT13 9FF, UK. victor.beaumont@pfizer.com., Liu L; Pfizer, Inc. BioTherapeutics Pharmaceutical Sciences, Analytical Research and Development, 1 Burtt Road, Andover, MA, 01810, USA., Shi H; Pfizer, Inc. Global Product Development, Oncology & Rare Disease Statistics, New York City, NY, 10001, USA., Rouse JC; Pfizer, Inc. BioTherapeutics Pharmaceutical Sciences, Analytical Research and Development, 1 Burtt Road, Andover, MA, 01810, USA., Kim HY; Pfizer, Inc. BioTherapeutics Pharmaceutical Sciences, Analytical Research and Development, 1 Burtt Road, Andover, MA, 01810, USA. hai-young.kim@pfizer.com.
Jazyk: angličtina
Zdroj: Pharmaceutical research [Pharm Res] 2023 Oct; Vol. 40 (10), pp. 2457-2467. Date of Electronic Publication: 2023 Oct 05.
DOI: 10.1007/s11095-023-03600-2
Abstrakt: Purpose: Nuclear magnetic resonance (NMR) spectroscopy provides the sensitivity and specificity to probe the higher order structure (HOS) of monoclonal antibodies (mAbs) for potential changes. This study demonstrates an application of chemometric tools to measure differences in the NMR spectra of mAbs after forced degradation relative to the respective unstressed starting materials.
Methods: Samples of adalimumab (Humira, ADL-REF) and trastuzumab (Herceptin, TRA-REF) were incubated in three buffer-pH conditions at 40°C for 4 weeks to compare to a control sample that was left unstressed. Replicate 1D 1 H and 2D 1 H- 13 C HMQC NMR spectra were collected on all samples. Chemometric analyses such as Easy Comparability of HOS (ECHOS), PROtein FIngerprinting by Lineshape Enhancement (PROFILE), and Principal Component Analysis (PCA) were applied to capture and quantitate differences between the spectra.
Results: Visual and statistical inspection of the 2D 1 H- 13 C HMQC spectra of adalimumab and trastuzumab after forced degradation conditions shows no changes in the spectra relative to the unstressed material. Chemometric analysis of the 1D 1 H NMR spectra shows only minor changes in the spectra of adalimumab after forced degradation, but significant differences in trastuzumab.
Conclusion: The chemometric analyses support the lack of statistical differences in the structure of pH-thermal stressed adalimumab, however, it reveals conformational changes or chemical modifications in trastuzumab after forced degradation. Application of chemometrics in comparative NMR studies enables HOS characterization and showcases the sensitivity and specificity in detecting differences in the spectra of mAbs after pH-thermal forced degradation with respect to local and global protein structure.
(© 2023. The Author(s).)
Databáze: MEDLINE
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