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On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS)

The crystallization of Anti-CD20, a full-length monoclonal antibody, has been studied in the PEG400/Na(2)SO(4)/Water system near Liquid-Liquid Phase Separation (LLPS) conditions by both sitting-drop vapour diffusion and batch methods. In order to understand the Anti-CD20 crystallization propensity i...

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Autores principales: Pantuso, Elvira, Mastropietro, Teresa F., Briuglia, Maria L., Gerard, Charline J. J., Curcio, Efrem, ter Horst, Joop H., Nicoletta, Fiore P., Di Profio, Gianluca
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264149/
https://www.ncbi.nlm.nih.gov/pubmed/32483267
http://dx.doi.org/10.1038/s41598-020-65776-6
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author Pantuso, Elvira
Mastropietro, Teresa F.
Briuglia, Maria L.
Gerard, Charline J. J.
Curcio, Efrem
ter Horst, Joop H.
Nicoletta, Fiore P.
Di Profio, Gianluca
author_facet Pantuso, Elvira
Mastropietro, Teresa F.
Briuglia, Maria L.
Gerard, Charline J. J.
Curcio, Efrem
ter Horst, Joop H.
Nicoletta, Fiore P.
Di Profio, Gianluca
author_sort Pantuso, Elvira
collection PubMed
description The crystallization of Anti-CD20, a full-length monoclonal antibody, has been studied in the PEG400/Na(2)SO(4)/Water system near Liquid-Liquid Phase Separation (LLPS) conditions by both sitting-drop vapour diffusion and batch methods. In order to understand the Anti-CD20 crystallization propensity in the solvent system of different compositions, we investigated some measurable parameters, normally used to assess protein conformational and colloidal stability in solution, with the aim to understand the aggregation mechanism of this complex biomacromolecule. We propose that under crystallization conditions a minor population of specifically aggregated protein molecules are present. While this minor species hardly contributes to the measured average solution behaviour, it induces and promotes crystal formation. The existence of this minor species is the result of the LLPS occurring concomitantly under crystallization conditions.
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spelling pubmed-72641492020-06-05 On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS) Pantuso, Elvira Mastropietro, Teresa F. Briuglia, Maria L. Gerard, Charline J. J. Curcio, Efrem ter Horst, Joop H. Nicoletta, Fiore P. Di Profio, Gianluca Sci Rep Article The crystallization of Anti-CD20, a full-length monoclonal antibody, has been studied in the PEG400/Na(2)SO(4)/Water system near Liquid-Liquid Phase Separation (LLPS) conditions by both sitting-drop vapour diffusion and batch methods. In order to understand the Anti-CD20 crystallization propensity in the solvent system of different compositions, we investigated some measurable parameters, normally used to assess protein conformational and colloidal stability in solution, with the aim to understand the aggregation mechanism of this complex biomacromolecule. We propose that under crystallization conditions a minor population of specifically aggregated protein molecules are present. While this minor species hardly contributes to the measured average solution behaviour, it induces and promotes crystal formation. The existence of this minor species is the result of the LLPS occurring concomitantly under crystallization conditions. Nature Publishing Group UK 2020-06-01 /pmc/articles/PMC7264149/ /pubmed/32483267 http://dx.doi.org/10.1038/s41598-020-65776-6 Text en © The Author(s) 2020 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Pantuso, Elvira
Mastropietro, Teresa F.
Briuglia, Maria L.
Gerard, Charline J. J.
Curcio, Efrem
ter Horst, Joop H.
Nicoletta, Fiore P.
Di Profio, Gianluca
On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS)
title On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS)
title_full On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS)
title_fullStr On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS)
title_full_unstemmed On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS)
title_short On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS)
title_sort on the aggregation and nucleation mechanism of the monoclonal antibody anti-cd20 near liquid-liquid phase separation (llps)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264149/
https://www.ncbi.nlm.nih.gov/pubmed/32483267
http://dx.doi.org/10.1038/s41598-020-65776-6
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