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Use of the 2D (1)H-(13)C HSQC NMR Methyl Region to Evaluate the Higher Order Structural Integrity of Biopharmaceuticals

The higher-order structure (HOS) of protein therapeutics is directly related to the function and represents a critical quality attribute. Currently, the HOS of protein therapeutics is characterized by methods with low to medium structural resolution, such as Fourier transform infrared (FTIR), circul...

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Autores principales: Hwang, Tsang-Lin, Batabyal, Dipanwita, Knutson, Nicholas, Wikström, Mats
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125315/
https://www.ncbi.nlm.nih.gov/pubmed/34063095
http://dx.doi.org/10.3390/molecules26092714
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author Hwang, Tsang-Lin
Batabyal, Dipanwita
Knutson, Nicholas
Wikström, Mats
author_facet Hwang, Tsang-Lin
Batabyal, Dipanwita
Knutson, Nicholas
Wikström, Mats
author_sort Hwang, Tsang-Lin
collection PubMed
description The higher-order structure (HOS) of protein therapeutics is directly related to the function and represents a critical quality attribute. Currently, the HOS of protein therapeutics is characterized by methods with low to medium structural resolution, such as Fourier transform infrared (FTIR), circular dichroism (CD), intrinsic fluorescence spectroscopy (FLD), and differential scanning calorimetry (DSC). High-resolution nuclear magnetic resonance (NMR) methods have now been introduced, representing powerful approaches for HOS characterization (HOS by NMR). NMR is a multi-attribute method with unique abilities to give information on all structural levels of proteins in solution. In this study, we have compared 2D (1)H-(13)C HSQC NMR with two established biophysical methods, i.e., near-ultraviolet circular dichroism (NUV-CD) and intrinsic fluorescence spectroscopy, for the HOS assessments for the folded and unfolded states of two monoclonal antibodies belonging to the subclasses IgG1 and IgG2. The study shows that the methyl region of the (1)H-(13)C HSQC NMR spectrum is sensitive to both the secondary and tertiary structure of proteins and therefore represents a powerful tool in assessing the overall higher-order structural integrity of biopharmaceutical molecules.
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spelling pubmed-81253152021-05-17 Use of the 2D (1)H-(13)C HSQC NMR Methyl Region to Evaluate the Higher Order Structural Integrity of Biopharmaceuticals Hwang, Tsang-Lin Batabyal, Dipanwita Knutson, Nicholas Wikström, Mats Molecules Article The higher-order structure (HOS) of protein therapeutics is directly related to the function and represents a critical quality attribute. Currently, the HOS of protein therapeutics is characterized by methods with low to medium structural resolution, such as Fourier transform infrared (FTIR), circular dichroism (CD), intrinsic fluorescence spectroscopy (FLD), and differential scanning calorimetry (DSC). High-resolution nuclear magnetic resonance (NMR) methods have now been introduced, representing powerful approaches for HOS characterization (HOS by NMR). NMR is a multi-attribute method with unique abilities to give information on all structural levels of proteins in solution. In this study, we have compared 2D (1)H-(13)C HSQC NMR with two established biophysical methods, i.e., near-ultraviolet circular dichroism (NUV-CD) and intrinsic fluorescence spectroscopy, for the HOS assessments for the folded and unfolded states of two monoclonal antibodies belonging to the subclasses IgG1 and IgG2. The study shows that the methyl region of the (1)H-(13)C HSQC NMR spectrum is sensitive to both the secondary and tertiary structure of proteins and therefore represents a powerful tool in assessing the overall higher-order structural integrity of biopharmaceutical molecules. MDPI 2021-05-05 /pmc/articles/PMC8125315/ /pubmed/34063095 http://dx.doi.org/10.3390/molecules26092714 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hwang, Tsang-Lin
Batabyal, Dipanwita
Knutson, Nicholas
Wikström, Mats
Use of the 2D (1)H-(13)C HSQC NMR Methyl Region to Evaluate the Higher Order Structural Integrity of Biopharmaceuticals
title Use of the 2D (1)H-(13)C HSQC NMR Methyl Region to Evaluate the Higher Order Structural Integrity of Biopharmaceuticals
title_full Use of the 2D (1)H-(13)C HSQC NMR Methyl Region to Evaluate the Higher Order Structural Integrity of Biopharmaceuticals
title_fullStr Use of the 2D (1)H-(13)C HSQC NMR Methyl Region to Evaluate the Higher Order Structural Integrity of Biopharmaceuticals
title_full_unstemmed Use of the 2D (1)H-(13)C HSQC NMR Methyl Region to Evaluate the Higher Order Structural Integrity of Biopharmaceuticals
title_short Use of the 2D (1)H-(13)C HSQC NMR Methyl Region to Evaluate the Higher Order Structural Integrity of Biopharmaceuticals
title_sort use of the 2d (1)h-(13)c hsqc nmr methyl region to evaluate the higher order structural integrity of biopharmaceuticals
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125315/
https://www.ncbi.nlm.nih.gov/pubmed/34063095
http://dx.doi.org/10.3390/molecules26092714
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