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...
Main Authors: | , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Nature Publishing Group
2021-04-01
|
Series: | Scientific Reports |
Online Access: | https://doi.org/10.1038/s41598-021-87912-6 |
id |
doaj-1707556cd31c43a4a60f3a8bae3edeac |
---|---|
record_format |
Article |
spelling |
doaj-1707556cd31c43a4a60f3a8bae3edeac2021-04-18T11:39:25ZengNature Publishing GroupScientific Reports2045-23222021-04-0111111110.1038/s41598-021-87912-6A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cellsYuki Noguchi0Kazuhisa Ozeki1Hiroaki Takesue2Hidetaka Akita3Research Division, Chugai Pharmaceutical Co., Ltd.Research Division, Chugai Pharmaceutical Co., Ltd.Research Division, Chugai Pharmaceutical Co., Ltd.Laboratory of DDS Design and Drug Disposition, Graduate School of Pharmaceutical Sciences, Chiba UniversityAbstract 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.https://doi.org/10.1038/s41598-021-87912-6 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yuki Noguchi Kazuhisa Ozeki Hiroaki Takesue Hidetaka Akita |
spellingShingle |
Yuki Noguchi Kazuhisa Ozeki Hiroaki Takesue Hidetaka Akita A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells Scientific Reports |
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 Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2021-04-01 |
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. |
url |
https://doi.org/10.1038/s41598-021-87912-6 |
work_keys_str_mv |
AT yukinoguchi acellbasedassayforevaluatingbindinganduptakeofanantibodyusinghepaticnonparenchymalcells AT kazuhisaozeki acellbasedassayforevaluatingbindinganduptakeofanantibodyusinghepaticnonparenchymalcells AT hiroakitakesue acellbasedassayforevaluatingbindinganduptakeofanantibodyusinghepaticnonparenchymalcells AT hidetakaakita acellbasedassayforevaluatingbindinganduptakeofanantibodyusinghepaticnonparenchymalcells AT yukinoguchi cellbasedassayforevaluatingbindinganduptakeofanantibodyusinghepaticnonparenchymalcells AT kazuhisaozeki cellbasedassayforevaluatingbindinganduptakeofanantibodyusinghepaticnonparenchymalcells AT hiroakitakesue cellbasedassayforevaluatingbindinganduptakeofanantibodyusinghepaticnonparenchymalcells AT hidetakaakita cellbasedassayforevaluatingbindinganduptakeofanantibodyusinghepaticnonparenchymalcells |
_version_ |
1721522010501152768 |