Purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles

Abstract Here, we describe porous zirconia particles (PZPs) optimized for the purification of immunoglobulins. PZPs, with a pore size of approximately 10 nm, were designed to specifically interact with antibodies via surface modification with a phosphate functional group. A simple PZP purification m...

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Autores principales:	Tetsuya Okuda, Katsuya Kato, Masahiro Kitamura, Shinjiro Kasahara
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b06165e97e294df58e0ddaef1ff40cd7
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spelling	oai:doaj.org-article:b06165e97e294df58e0ddaef1ff40cd72021-12-02T12:09:45ZPurification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles10.1038/s41598-021-82457-02045-2322https://doaj.org/article/b06165e97e294df58e0ddaef1ff40cd72021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82457-0https://doaj.org/toc/2045-2322Abstract Here, we describe porous zirconia particles (PZPs) optimized for the purification of immunoglobulins. PZPs, with a pore size of approximately 10 nm, were designed to specifically interact with antibodies via surface modification with a phosphate functional group. A simple PZP purification method based on precipitation enabled efficient purification of mouse anti-glycosphingolipid globoside/Gb4Cer monoclonal IgM (κ-light chains) from hybridoma culture supernatants. Over 99% of contaminating proteins were removed by the PZP purification process, and approximately 50% of the IgM was recovered in the purified fraction after eluting the PZP-adsorbed antibodies with 100 mM phosphate buffer. Other IgG3 and IgM monoclonal antibodies that react with Gb4Cer or α2,6-sialyl LacNAc-modified glycoproteins could also be purified using PZPs and elution buffer at concentrations of 100–500 mM. All of the purified antibodies retained their antigen reactivity and specificity, indicating that PZP purification does not affect antibody function. As PZP purification is also suitable for purification of IgM consisting of λ-light chains and IgG derived from other mammalian species, it is expected to be applied to the purification of a variety of antibodies, including anti-glycoconjugate IgMs.Tetsuya OkudaKatsuya KatoMasahiro KitamuraShinjiro KasaharaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tetsuya Okuda Katsuya Kato Masahiro Kitamura Shinjiro Kasahara Purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles
description	Abstract Here, we describe porous zirconia particles (PZPs) optimized for the purification of immunoglobulins. PZPs, with a pore size of approximately 10 nm, were designed to specifically interact with antibodies via surface modification with a phosphate functional group. A simple PZP purification method based on precipitation enabled efficient purification of mouse anti-glycosphingolipid globoside/Gb4Cer monoclonal IgM (κ-light chains) from hybridoma culture supernatants. Over 99% of contaminating proteins were removed by the PZP purification process, and approximately 50% of the IgM was recovered in the purified fraction after eluting the PZP-adsorbed antibodies with 100 mM phosphate buffer. Other IgG3 and IgM monoclonal antibodies that react with Gb4Cer or α2,6-sialyl LacNAc-modified glycoproteins could also be purified using PZPs and elution buffer at concentrations of 100–500 mM. All of the purified antibodies retained their antigen reactivity and specificity, indicating that PZP purification does not affect antibody function. As PZP purification is also suitable for purification of IgM consisting of λ-light chains and IgG derived from other mammalian species, it is expected to be applied to the purification of a variety of antibodies, including anti-glycoconjugate IgMs.
format	article
author	Tetsuya Okuda Katsuya Kato Masahiro Kitamura Shinjiro Kasahara
author_facet	Tetsuya Okuda Katsuya Kato Masahiro Kitamura Shinjiro Kasahara
author_sort	Tetsuya Okuda
title	Purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles
title_short	Purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles
title_full	Purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles
title_fullStr	Purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles
title_full_unstemmed	Purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles
title_sort	purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b06165e97e294df58e0ddaef1ff40cd7
work_keys_str_mv	AT tetsuyaokuda purificationofantiglycoconjugatemonoclonalantibodiesusingnewlydevelopedporouszirconiaparticles AT katsuyakato purificationofantiglycoconjugatemonoclonalantibodiesusingnewlydevelopedporouszirconiaparticles AT masahirokitamura purificationofantiglycoconjugatemonoclonalantibodiesusingnewlydevelopedporouszirconiaparticles AT shinjirokasahara purificationofantiglycoconjugatemonoclonalantibodiesusingnewlydevelopedporouszirconiaparticles
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Purification of anti-glycoconjugate monoclonal antibodies using newly developed porous zirconia particles

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