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

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Title: A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells.
Authors: Noguchi, Yuki, Ozeki, Kazuhisa, Takesue, Hiroaki, Akita, Hidetaka
Source: Scientific Reports; 4/16/2021, Vol. 11 Issue 1, p1-11, 11p
Subject Terms: IMMUNOGLOBULIN G receptors, ENDOTHELIAL cells, PHARMACOKINETICS, FLOW cytometry, BIOLOGICALS, LIVER 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 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. [ABSTRACT FROM AUTHOR]
Copyright of Scientific Reports is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells.
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  Data: <searchLink fieldCode="AR" term="%22Noguchi%2C+Yuki%22">Noguchi, Yuki</searchLink><br /><searchLink fieldCode="AR" term="%22Ozeki%2C+Kazuhisa%22">Ozeki, Kazuhisa</searchLink><br /><searchLink fieldCode="AR" term="%22Takesue%2C+Hiroaki%22">Takesue, Hiroaki</searchLink><br /><searchLink fieldCode="AR" term="%22Akita%2C+Hidetaka%22">Akita, Hidetaka</searchLink>
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  Data: Scientific Reports; 4/16/2021, Vol. 11 Issue 1, p1-11, 11p
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  Data: <searchLink fieldCode="DE" term="%22IMMUNOGLOBULIN+G+receptors%22">IMMUNOGLOBULIN G receptors</searchLink><br /><searchLink fieldCode="DE" term="%22ENDOTHELIAL+cells%22">ENDOTHELIAL cells</searchLink><br /><searchLink fieldCode="DE" term="%22PHARMACOKINETICS%22">PHARMACOKINETICS</searchLink><br /><searchLink fieldCode="DE" term="%22FLOW+cytometry%22">FLOW cytometry</searchLink><br /><searchLink fieldCode="DE" term="%22BIOLOGICALS%22">BIOLOGICALS</searchLink><br /><searchLink fieldCode="DE" term="%22LIVER+cells%22">LIVER cells</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: 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. [ABSTRACT FROM AUTHOR]
– Name: Abstract
  Label:
  Group: Ab
  Data: <i>Copyright of Scientific Reports is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1038/s41598-021-87912-6
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        Text: English
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              Text: 4/16/2021
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              Y: 2021
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