2D 1HN, 15N Correlated NMR Methods at Natural Abundance for Obtaining Structural Maps and Statistical Comparability of Monoclonal Antibodies

Published

March 1, 2015

Author(s)

Luke Arbogast, Robert G. Brinson, Catherine A. Mouchahoir, James T. Hoopes, John Marino

Abstract

Purpose High-resolution nuclear magnetic resonance spectroscopy (NMR) provides a robust approach for producing unique spectral signatures of protein higher order structure at atomic resolution. Such signatures can be used as a tool to establish consistency of protein folding for the assessment of monoclonal antibody (mAb) drug quality and comparability. Methods Using the NIST monoclonal antibody (NISTmAb) and a commercial-sourced polyclonal antibody, both IgG1κ isotype, we apply 2D NMR methods at natural abundance for the acquisition and unbiased statistical analysis of 1HN -15N correlated spectra of intact antibody (Ab) and protease- cleaved Fab and Fc fragments. Results The study demonstrates the feasibility of applying 2D NMR techniques to Abs and the precision with which these methods can be used to map structure and establish comparability between samples at atomic resolution. Conclusions The statistical analyses suggests that, within the limit of detection, no significant structural differences are observed between the Fab and Fc domains of each respective intact Ab and its corresponding fragments. Discrimination between dissimilar species, such as between the Fab domains of both Abs or between the glycosylated and deglycosylated Fc domains, was further demonstrated. As such, these methods should find general utility for the assessment of mAb higher order structure.

Citation

Pharmaceutical Research

Volume

87

Issue

7

Pub Type

Journals

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DOI Link

Keywords

high resolution NMR, higher order structure, biosimilars

Citation

Arbogast, L. , Brinson, R. , Mouchahoir, C. , Hoopes, J. and Marino, J. (2015), 2D 1HN, 15N Correlated NMR Methods at Natural Abundance for Obtaining Structural Maps and Statistical Comparability of Monoclonal Antibodies, Pharmaceutical Research, [online], https://doi.org/10.1021/ac504804m (Accessed April 18, 2024)