A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells

Abstract Evaluation of the binding and uptake of an antibody in liver non-parenchymal cells (NPC), including liver sinusoidal endothelial cells, is important for revealing its pharmacokinetic (PK) behavior, since NPC has important roles in eliminating an antibody from the blood via the Fc fragment o...

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Autores principales:	Yuki Noguchi, Kazuhisa Ozeki, Hiroaki Takesue, Hidetaka Akita
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1707556cd31c43a4a60f3a8bae3edeac
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spelling	oai:doaj.org-article:1707556cd31c43a4a60f3a8bae3edeac2021-12-02T18:03:21ZA cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells10.1038/s41598-021-87912-62045-2322https://doaj.org/article/1707556cd31c43a4a60f3a8bae3edeac2021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-87912-6https://doaj.org/toc/2045-2322Abstract Evaluation of the binding and uptake of an antibody in liver non-parenchymal cells (NPC), including liver sinusoidal endothelial cells, is important for revealing its pharmacokinetic (PK) behavior, since NPC has important roles in eliminating an antibody from the blood via the Fc fragment of IgG receptor IIB (FcγRIIB). However, there is currently no in vitro quantitative assay using NPC. This study reports on the development of a cell-based assay for evaluating the binding and uptake of such an antibody using liver NPC of mice and monkeys. In mice, the FcγRIIB-expressing cells were identified in the CD146-positive and CD45-negative fraction by flow cytometry. A titration assay was performed to determine the PK parameters, and the obtained parameter was comparable to that determined by the fitting of the in vivo PK. This approach was also extended to NPC from monkeys. The concentration-dependent binding and uptake was measured to determine the PK parameters using monkey NPC, the FcγRIIB-expressing fraction of which was identified by CD31 and CD45. The findings presented herein demonstrate that the in vitro liver NPC assay using flow cytometry is a useful tool to determine the binding and uptake of biologics and to predict the PK.Yuki NoguchiKazuhisa OzekiHiroaki TakesueHidetaka AkitaNature 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 Yuki Noguchi Kazuhisa Ozeki Hiroaki Takesue Hidetaka Akita A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells
description	Abstract Evaluation of the binding and uptake of an antibody in liver non-parenchymal cells (NPC), including liver sinusoidal endothelial cells, is important for revealing its pharmacokinetic (PK) behavior, since NPC has important roles in eliminating an antibody from the blood via the Fc fragment of IgG receptor IIB (FcγRIIB). However, there is currently no in vitro quantitative assay using NPC. This study reports on the development of a cell-based assay for evaluating the binding and uptake of such an antibody using liver NPC of mice and monkeys. In mice, the FcγRIIB-expressing cells were identified in the CD146-positive and CD45-negative fraction by flow cytometry. A titration assay was performed to determine the PK parameters, and the obtained parameter was comparable to that determined by the fitting of the in vivo PK. This approach was also extended to NPC from monkeys. The concentration-dependent binding and uptake was measured to determine the PK parameters using monkey NPC, the FcγRIIB-expressing fraction of which was identified by CD31 and CD45. The findings presented herein demonstrate that the in vitro liver NPC assay using flow cytometry is a useful tool to determine the binding and uptake of biologics and to predict the PK.
format	article
author	Yuki Noguchi Kazuhisa Ozeki Hiroaki Takesue Hidetaka Akita
author_facet	Yuki Noguchi Kazuhisa Ozeki Hiroaki Takesue Hidetaka Akita
author_sort	Yuki Noguchi
title	A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells
title_short	A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells
title_full	A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells
title_fullStr	A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells
title_full_unstemmed	A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells
title_sort	cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1707556cd31c43a4a60f3a8bae3edeac
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A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells

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