Purification of antibody fragments via interaction with detergent micellar aggregates

Abstract The research described in this report seeks to present proof-of-concept for a novel and robust platform for purification of antibody fragments and to define and optimize the controlling parameters. Purification of antigen-binding F(ab′)2 fragments is achieved in the absence of chromatograph...

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Autores principales:	Gunasekaran Dhandapani, Ellen Wachtel, Ishita Das, Mordechai Sheves, Guy Patchornik
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e82c8facc746412893f993be950f9808
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spelling	oai:doaj.org-article:e82c8facc746412893f993be950f98082021-12-02T18:25:03ZPurification of antibody fragments via interaction with detergent micellar aggregates10.1038/s41598-021-90966-12045-2322https://doaj.org/article/e82c8facc746412893f993be950f98082021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90966-1https://doaj.org/toc/2045-2322Abstract The research described in this report seeks to present proof-of-concept for a novel and robust platform for purification of antibody fragments and to define and optimize the controlling parameters. Purification of antigen-binding F(ab′)2 fragments is achieved in the absence of chromatographic media or specific ligands, rather by using clusters of non-ionic detergent (e.g. Tween-60, Brij-O20) micelles chelated via Fe2+ ions and the hydrophobic chelator, bathophenanthroline (batho). These aggregates, quantitatively capture the F(ab′)2 fragment in the absence or presence of E. coli lysate and allow extraction of only the F(ab′)2 domain at pH 3.8 without concomitant aggregate dissolution or coextraction of bacterial impurities. Process yields range from 70 to 87% by densitometry. Recovered F(ab′)2 fragments are monomeric (by dynamic light scattering), preserve their secondary structure (by circular dichroism) and are as pure as those obtained via Protein A chromatography (from a mixture of F(ab′)2 and Fc fragments). The effect of process parameters on Ab binding and Ab extraction (e.g. temperature, pH, ionic strength, incubation time, composition of extraction buffer) are reported, using a monoclonal antibody (mAb) and polyclonal human IgG’s as test samples.Gunasekaran DhandapaniEllen WachtelIshita DasMordechai ShevesGuy PatchornikNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Gunasekaran Dhandapani Ellen Wachtel Ishita Das Mordechai Sheves Guy Patchornik Purification of antibody fragments via interaction with detergent micellar aggregates
description	Abstract The research described in this report seeks to present proof-of-concept for a novel and robust platform for purification of antibody fragments and to define and optimize the controlling parameters. Purification of antigen-binding F(ab′)2 fragments is achieved in the absence of chromatographic media or specific ligands, rather by using clusters of non-ionic detergent (e.g. Tween-60, Brij-O20) micelles chelated via Fe2+ ions and the hydrophobic chelator, bathophenanthroline (batho). These aggregates, quantitatively capture the F(ab′)2 fragment in the absence or presence of E. coli lysate and allow extraction of only the F(ab′)2 domain at pH 3.8 without concomitant aggregate dissolution or coextraction of bacterial impurities. Process yields range from 70 to 87% by densitometry. Recovered F(ab′)2 fragments are monomeric (by dynamic light scattering), preserve their secondary structure (by circular dichroism) and are as pure as those obtained via Protein A chromatography (from a mixture of F(ab′)2 and Fc fragments). The effect of process parameters on Ab binding and Ab extraction (e.g. temperature, pH, ionic strength, incubation time, composition of extraction buffer) are reported, using a monoclonal antibody (mAb) and polyclonal human IgG’s as test samples.
format	article
author	Gunasekaran Dhandapani Ellen Wachtel Ishita Das Mordechai Sheves Guy Patchornik
author_facet	Gunasekaran Dhandapani Ellen Wachtel Ishita Das Mordechai Sheves Guy Patchornik
author_sort	Gunasekaran Dhandapani
title	Purification of antibody fragments via interaction with detergent micellar aggregates
title_short	Purification of antibody fragments via interaction with detergent micellar aggregates
title_full	Purification of antibody fragments via interaction with detergent micellar aggregates
title_fullStr	Purification of antibody fragments via interaction with detergent micellar aggregates
title_full_unstemmed	Purification of antibody fragments via interaction with detergent micellar aggregates
title_sort	purification of antibody fragments via interaction with detergent micellar aggregates
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/e82c8facc746412893f993be950f9808
work_keys_str_mv	AT gunasekarandhandapani purificationofantibodyfragmentsviainteractionwithdetergentmicellaraggregates AT ellenwachtel purificationofantibodyfragmentsviainteractionwithdetergentmicellaraggregates AT ishitadas purificationofantibodyfragmentsviainteractionwithdetergentmicellaraggregates AT mordechaisheves purificationofantibodyfragmentsviainteractionwithdetergentmicellaraggregates AT guypatchornik purificationofantibodyfragmentsviainteractionwithdetergentmicellaraggregates
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Purification of antibody fragments via interaction with detergent micellar aggregates

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