Multivariate Analysis of 2D (1)H,(13)C methyl NMR Spectra of Monoclonal Antibody Therapeutics to Facilitate Assessment of Higher Order Structure.

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TitleMultivariate Analysis of 2D (1)H,(13)C methyl NMR Spectra of Monoclonal Antibody Therapeutics to Facilitate Assessment of Higher Order Structure.
Publication TypeJournal Article
Year of Publication2017
AuthorsArbogast, L, Delaglio, F, Schiel, JE, Marino, JP
JournalAnal Chem
Date Published2017 Sep 22
ISSN1520-6882
Abstract

Two-dimensional (2D) 1H-13C methyl NMR provides a powerful tool to probe the higher order structure (HOS) of monoclonal antibodies (mAbs), since spectra can readily be acquired on intact mAbs at natural isotopic abundance, and small changes in chemical environment and structure give rise to observable changes in corresponding spectra, which can be interpreted at atomic resolution. This makes it possible to apply 2D NMR spectral fingerprinting approaches directly to drug products in order to systematically characterize structure and excipient effects. Systematic collections of NMR spectra are often analyzed in terms of the changes in specifically identified peak positions, as well as changes in peak height and linewidths. A complementary approach is to apply Principal Component Analysis (PCA) directly to the matrix of spectral data, correlating spectra according to similarities and differences in their overall shapes, rather than according to parameters of individually identified peaks. This is particularly well-suited for spectra of mAbs, where some of the individual peaks might not be well resolved. Here we demonstrate the performance of the PCA method for discriminating structural variation among systematic sets of 2D NMR fingerprint spectra using the NISTmAb and illustrate how spectral variability identified by PCA may be correlated to structure.

DOI10.1021/acs.analchem.7b03571
Alternate JournalAnal. Chem.
PubMed ID28937210