Spin Diffusion Editing for Structural Fingerprints of Therapeutic Antibodies

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• 作者：Joshua Franks ; John N. Glushka ; Michael T. Jones ; David H. Live ; Qin Zou ; James H. Prestegard
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：January 19, 2016
• 年：2016
• 卷：88
• 期：2
• 页码：1320-1327
• 全文大小：422K
• ISSN：1520-6882

文摘

The growing importance of biologics and biosimilars as therapeutic and diagnostic agents is giving rise to new demands for analytical methodology that can quickly and accurately assess the chemical and physical state of protein-based products. A particular challenge exists in physical characterization where the proper fold and extent of disorder of a protein is a major concern. The ability of NMR to reflect structural and dynamic properties of proteins is well recognized, but sensitivity limitations and high levels of interference from excipients in typical biologic formulations have prevented widespread applications to quality assessment. Here we demonstrate applicability of a simple one-dimensional proton NMR method that exploits enhanced spin diffusion among protons in well-structured areas of a protein. We show that it is possible to reduce excipient signals and allow focus on structural characteristics of the protein. Additional decomposition of the resulting spectra based on rotating frame spin relaxation allows separate examination of components from aggregates and disordered regions. Application to a comparison of two different monoclonal antibodies and to detection of partial pH denaturation of a monoclonal antibody illustrates the procedure.