Detection and analysis of tupaia hepatocytes via mAbs against tupaia serum albumin

通过针对树鼩血清白蛋白的单克隆抗体检测和分析树鼩肝细胞

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作者:Xuan Liu, Lunzhi Yuan, Quan Yuan, Yali Zhang, Kun Wu, Tianying Zhang, Yong Wu, Wangheng Hou, Tengyun Wang, Pingguo Liu, James Wai Kuo Shih, Tong Cheng, Ningshao Xia

Abstract

On the basis of its close phylogenetic relationship with primates, the development of Tupaia belangeri as an infection animal model and drug metabolism model could provide a new option for preclinical studies, especially in hepatitis virus research. As a replacement for primary human hepatocytes (PHHs), primary tupaia hepatocytes (PTHs) have been widely used. Similar to human serum albumin, tupaia serum albumin (TSA) is the most common liver synthesis protein and is an important biomarker for PTHs and liver function. However, no detection or quantitative method for TSA has been reported. In this study, mouse monoclonal antibodies (mAbs) 4G5 and 9H3 against TSA were developed to recognize PTHs, and they did not show cross-reactivity with serum albumin from common experimental animals, such as the mouse, rat, cow, rabbit, goat, monkey, and chicken. The two mAbs also exhibited good performance in fluorescence activated cell sorting (FACS) analysis and immunofluorescence (IF) detection of PTHs. A chemiluminescent enzyme immune assay method using the two mAbs, with a linear range from 96.89 pg/ml to 49,609.38 pg/ml, was developed for the quantitative detection of TSA. The mAbs and the CLEIA method provide useful tools for research on TSA and PTHs.

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