A Simple and Low-cost Preliminary Quantification of Target Membrane Protein in Single Cells

doi:10.1007/s10895-023-03496-6

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	A Simple and Low-cost Preliminary Quantification of Target Membrane Protein in Single Cells
	Li, Jiamin 1,2,3; Shi, Meng 4; Geng, Xuhui 1,3; Guan, Yafeng 1,3,5
刊名	JOURNAL OF FLUORESCENCE
	2023-11-17
页码	8
关键词	Single cell Membrane Proteins Capillary Electrophoresis Fluorescence HER2 Protein
ISSN号	1053-0509
DOI	10.1007/s10895-023-03496-6
通讯作者	Guan, Yafeng(guanyafeng@dicp.ac.cn)
英文摘要	To study the heterogeneity of target membrane proteins in single cells with cellular integrity, we proposed a simple and low-cost method to obtain the copy number of the membrane proteins. HeLa cells were labeled by FITC affinity bodies specifically targeting HER2 membrane proteins. The immunolabeled HeLa cells were quantified by a laboratory-built laser induced fluorescence detector. A series of fluorescent microspheres with known number of FITC molecules on the surface were used to establish the calibration curve, instead of the standard fluorescent solutions, because the morphology of the microspheres was similar to the cells, and the distribution of FITC on the spheres were similar to the distribution of HER2 on the HeLa. The fluorescence intensity of the cells was converted to the molecule number of HER2 by the calibration curve. A capillary electrophoresis system was used to drive the microspheres and cells through the detection window. The copy number of HER2 in HeLa cells ranged from 4,036 to 1,224,920 +/- 100 (2.5-97.5%), and the median of copy numbers were 104,438 +/- 100 per cell. This method for measuring low-abundance membrane proteins can be utilized for the initial exploration of proteomics in ordinary laboratories.
资助项目	Thanks for professor Liming Wang of Dalian Medical University for the supplement of cells. Thanks for associated professor Shenghong Li of Dalian Institute of Chemical Physics for her effective suggestions.
WOS关键词	CAPILLARY-ELECTROPHORESIS ; COPY NUMBER ; MITOCHONDRIA ; EXPRESSION ; MOLECULES ; HER2
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
语种	英语
出版者	SPRINGER/PLENUM PUBLISHERS
WOS记录号	WOS:001105197900004
资助机构	Thanks for professor Liming Wang of Dalian Medical University for the supplement of cells. Thanks for associated professor Shenghong Li of Dalian Institute of Chemical Physics for her effective suggestions.
内容类型	期刊论文
源URL	[http://ir.idsse.ac.cn/handle/183446/10673]
专题	研究生部
通讯作者	Guan, Yafeng
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Dept Instrumentat & Analyt Chem, 457 Zhongshan Rd, Dalian 116023, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Deep Sea Composit Detect Technol Liaoning, Dalian 116023, Peoples R China 4.Liaoning Normal Univ, Sch Chem & Chem Engn, Dalian 116029, Peoples R China 5.Chinese Acad Sci, Inst Deep Sea Sci & Engn, 28 Luhuitou Rd, Sanya 572000, Peoples R China
推荐引用方式 GB/T 7714	Li, Jiamin,Shi, Meng,Geng, Xuhui,et al. A Simple and Low-cost Preliminary Quantification of Target Membrane Protein in Single Cells[J]. JOURNAL OF FLUORESCENCE,2023:8.
APA	Li, Jiamin,Shi, Meng,Geng, Xuhui,&Guan, Yafeng.(2023).A Simple and Low-cost Preliminary Quantification of Target Membrane Protein in Single Cells.JOURNAL OF FLUORESCENCE,8.
MLA	Li, Jiamin,et al."A Simple and Low-cost Preliminary Quantification of Target Membrane Protein in Single Cells".JOURNAL OF FLUORESCENCE (2023):8.