On the aggregation and nucleation mechanism of the monoclonal antibody anti-CD20 near liquid-liquid phase separation (LLPS)

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Pantuso, Elvira and Mastropietro, Teresa F. and Briuglia, Maria L. and Gerard, Charline J. J. and Curcio, Efrem and ter Horst, Joop H. and Nicoletta, Fiore P. and Di Profio, Gianluca (2020) On the aggregation and nucleation mechanism of the monoclonal antibody anti-CD20 near liquid-liquid phase separation (LLPS). Scientific Reports, 10 (1). 8902. ISSN 2045-2322 (https://doi.org/10.1038/s41598-020-65776-6)

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Abstract

The crystallization of Anti-CD20, a full-length monoclonal antibody, has been studied in the PEG400/Na2SO4/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.

ORCID iDs

Pantuso, Elvira, Mastropietro, Teresa F., Briuglia, Maria L. ORCID logoORCID: https://orcid.org/0000-0002-1737-0767, Gerard, Charline J. J., Curcio, Efrem, ter Horst, Joop H. ORCID logoORCID: https://orcid.org/0000-0003-0118-2160, Nicoletta, Fiore P. and Di Profio, Gianluca;
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