Development of a nanobody-based immunoassay for the sensitive detection of fibrinogen-like protein 1

doi:10.1038/s41401-020-00574-4

	Development of a nanobody-based immunoassay for the sensitive detection of fibrinogen-like protein 1
	Zhang, Wan-ting 5; Liu, Ting-ting 6; Wu, Man 7; Chen, Xiao-chen 6; Han, Li 7; Shi, Zhen-zhong 2; Li, Yu-ying 3,6; Li, Xi-yang 4; Xu, Hai-xing 5; Gong, Li-kun 1,3,6
刊名	ACTA PHARMACOLOGICA SINICA
	2021-02-25
页码	9
关键词	nanobody FGL1 cancer immunology diagnosis immune checkpoint
ISSN号	1671-4083
DOI	10.1038/s41401-020-00574-4
通讯作者	Xu, Hai-xing(3186@whut.edu.cn) ; Gong, Li-kun(lkgong@cdser.simm.ac.cn) ; Xu, Pei-hu(whutxph68@126.com) ; Geng, Yong(gengyong@simm.ac.cn)
英文摘要	Immune checkpoint inhibition is an important strategy in cancer therapy. Blockade of CTLA-4 and PD-1/PD-L1 is well developed in clinical practice. In the last few years, LAG-3 has received much interest as an emerging novel target in immunotherapy. It was recently reported that FGL1 is a major ligand of LAG-3, which is normally secreted by the liver but is upregulated in several human cancers. FGL1 is a crucial biomarker and target for cancer immunotherapy. As the efficacy of immunotherapy is limited to specific types of patients, the subset of patients needs to be selected appropriately to receive precise treatment according to different biomarkers. To date, there is no test to accurately assess FGL1 expression levels. Nanobodies have some outstanding features, such as high stability, solubility and affinity for diagnostic and therapeutic applications. Here, we report the development and validation of a rapid, sensitive, and cost-effective nanobody-based immunoassay for the detection of FGL1 in human serum. In this study, human FGL1 recombinant protein was expressed and purified for the first time as an immunized antigen. Then, we constructed a nanobody phage display library and screened several nanobodies that bind FGL1 with high affinity. We selected two nanobodies targeting different epitopes of FGL1, one as a capture and the other conjugated with HRP as a probe. The double nanobody-based sandwich ELISA to detect the concentration of FGL1 showed a good response relationship in the range of 15.625-2000 ng/mL, and the recoveries from the spiked sample were in the range of 78% and 100%. This assay could be used as a potential approach for evaluating FGL1 expression for patient stratification and for predicting the therapeutic efficacy of targeting the LAG3/FGL1 axis.
资助项目	National Natural Science Foundation of China[31670743] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDA12040326] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDA12050305] ; Science and Technology Commission of Shanghai Municipality[3918JC141540001] ; Joint Research Fund for Overseas, Hong Kong and Macao Scholars[81628013] ; National New Drug Creation Program of China[2018ZX09201017-004] ; National New Drug Creation Program of China[2019ZX09732002-013] ; Natural Science Foundation of Shanghai[16ZR1442900] ; National Science Foundation for Young Scholar projects[118180359901] ; Shanghai Institute of Materia Medica ; Chinese Academy of Sciences[CASIMM0120164013] ; Chinese Academy of Sciences[SIMM1606YZZ-06] ; Chinese Academy of Sciences[SIMM1601KF-06] ; Chinese Academy of Sciences[55201631121116101] ; Chinese Academy of Sciences[55201631121108000] ; Chinese Academy of Sciences[5112345601] ; Chinese Academy of Sciences[2015123456005]
WOS研究方向	Chemistry ; Pharmacology & Pharmacy
语种	英语
出版者	NATURE PUBLISHING GROUP
WOS记录号	WOS:000621723400002
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/295641]
专题	中国科学院上海药物研究所
通讯作者	Xu, Hai-xing; Gong, Li-kun; Xu, Pei-hu; Geng, Yong
作者单位	1.Chinese Acad Sci, Zhongshan Inst Drug Discovery, Inst Drug Discovery & Dev, Zhongshan 528400, Peoples R China 2.Shanghai Ocean Univ, Coll Food Sci & Technol, Shanghai 201306, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Nanjing Univ Chinese Med, Sch Novel Chinese Med, Nanjing 210046, Peoples R China 5.Wuhan Univ Technol, Sch Chem Chem Engn & Life Sci, Wuhan 420070, Peoples R China 6.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 7.Shanghai Univ Tradit Chinese Med, Sch Pharm, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Wan-ting,Liu, Ting-ting,Wu, Man,et al. Development of a nanobody-based immunoassay for the sensitive detection of fibrinogen-like protein 1[J]. ACTA PHARMACOLOGICA SINICA,2021:9.
APA	Zhang, Wan-ting.,Liu, Ting-ting.,Wu, Man.,Chen, Xiao-chen.,Han, Li.,...&Geng, Yong.(2021).Development of a nanobody-based immunoassay for the sensitive detection of fibrinogen-like protein 1.ACTA PHARMACOLOGICA SINICA,9.
MLA	Zhang, Wan-ting,et al."Development of a nanobody-based immunoassay for the sensitive detection of fibrinogen-like protein 1".ACTA PHARMACOLOGICA SINICA (2021):9.