Principal Component Analysis of 1D ¹H Diffusion Edited NMR Spectra of Protein Therapeutics.

Published

June 20, 2021

Author(s)

Korth Elliott, Frank Delaglio, Mats Wikstrom, John Marino, Luke Arbogast

Abstract

The one-dimensional (1D) diffusion edited proton NMR method, Protein Fingerprint by Lineshape Enhancement (PROFILE) has been demonstrated to be suitable for higher order structure (HOS) characterization of protein therapeutics including monoclonal antibodies. Recent reports in the literature have demonstrated its advantages for HOS characterization over traditional methods such as circular dichroism and Fourier-transform infrared spectroscopy. Previously, we have demonstrated that the PROFILE method is complementary with high resolution 2D methyl correlated NMR methods and how both may be deployed as a multi-modal platform to further the utility of NMR for HOS characterization. A major limitation of the PROFILE method remains its need for high signal to noise data due to its reliance on convolution difference processing and linear correlation metrics to assess spectral similarity. Here we present an alternative method for analyzing 1D diffusion edited spectra, which overcomes this limitation by using nonlinear iterative partial least squares (NIPALS) principal component analysis, and which we dub PROtein Fingerprint Observed Using NIPALS Decomposition (PROFOUND). We demonstrate that results from the PROFOUND method are robust with respect to instrument, operator and in the presence of high experimental noise and how it may be employed to provide quantitative assessment of spectral similarity.

Citation

Journal of Pharmaceutical Sciences

Pub Type

Journals

Download Paper

https://doi.org/10.1016/j.xphs.2021.06.027

Local Download

Keywords

Nuclear Magnetic Resonance (NMR), spectroscopy monoclonal antibody(s), Principal component analysis (PCA), Protein structure(s), Chemometrics, PROFOUND

Citation

Elliott, K. , Delaglio, F. , Wikstrom, M. , Marino, J. and Arbogast, L. (2021), Principal Component Analysis of 1D ¹H Diffusion Edited NMR Spectra of Protein Therapeutics., Journal of Pharmaceutical Sciences, [online], https://doi.org/10.1016/j.xphs.2021.06.027, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=931898 (Accessed October 8, 2025)