Quantification of Low Copy Number Proteins in Single Cells
文献类型：期刊论文
| 作者 | Shi, Meng1,2; Geng, Xuhui1; Wang, Chengye3; Guan, Yafeng1 |
| 刊名 | ANALYTICAL CHEMISTRY
 |
| 出版日期 | 2019-09-17 |
| 卷号 | 91期号:18页码:11493-11496 |
| ISSN号 | 0003-2700 |
| DOI | 10.1021/acs.analchem.9b02989 |
| 通讯作者 | Geng, Xuhui(xuhuigeng@126.com) ; Guan, Yafeng(guanyafeng@dicp.ac.cn) |
| 英文摘要 | We have developed an ultrasensitive and highly selective method to quantify low copy number intracellular proteins in a single cell using a low-cost laser-induced fluorescence (LIF) detector and a BV605 fluorescent probe. Active caspase3 proteins in cells were labeled by corresponding antibody-BV605 fluorescent binding, and a cell was injected into a 20 cm x 50 mu m i.d. capillary column, followed by in situ lysis and capillary electrophoresis (CE)-LIF analysis. About seven active caspase3 protein molecules in a detection volume of 91 pL could be detected. In our method, cross-bounding proteins other than active caspase3 could be separated and distinguished by differences of retention time. By using Si photodiode assembly as a fluorescent detector instead of PMT, the dynamic range of the LIF is over 4 orders of magnitude. In this experiment, we found that the number of active caspase3 molecules in 98 single Jurkat cells were from 629 to 12171, reflecting significant heterogeneity among the cells although they were from the same batch. For extended application, it could also be applied to quantify other types of low copy number proteins in a single cell as long as the corresponding antibodies are provided. This high-sensitive method could also be a promising tool for earlier cancer diagnosis and related disease pathway research which is relevant to low copy number proteins. In addition, this low-cost system could also be easily expanded to an array system for high-throughput quantitation of low copy proteins in single cells. |
| WOS关键词 | LASER-INDUCED FLUORESCENCE ; CAPILLARY-ELECTROPHORESIS ; SENSITIVITY ; SYSTEM ; EXPRESSION ; CASPASES ; DETECTOR |
| 资助项目 | DICP[DICP ZZBS201609] ; General Program of the National Natural Science Foundation of China[21874135] |
| WOS研究方向 | Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:000487156900002 |
| 出版者 | AMER CHEMICAL SOC |
| 资助机构 | DICP ; DICP ; General Program of the National Natural Science Foundation of China ; General Program of the National Natural Science Foundation of China ; DICP ; DICP ; General Program of the National Natural Science Foundation of China ; General Program of the National Natural Science Foundation of China ; DICP ; DICP ; General Program of the National Natural Science Foundation of China ; General Program of the National Natural Science Foundation of China ; DICP ; DICP ; General Program of the National Natural Science Foundation of China ; General Program of the National Natural Science Foundation of China |
| 源URL | [http://cas-ir.dicp.ac.cn/handle/321008/172797]  |
| 专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
| 通讯作者 | Geng, Xuhui; Guan, Yafeng |
| 作者单位 | 1.Chinese Acad Sci, Dalian Inst Chem Phys, Dept Instrumentat & Analyt Chem, CAS Key Lab Separat Sci Analyt Chem, 457 Zhongshan Rd, Dalian 116023, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Dalian Med Univ, Hosp 2, Dept Thorac Surg, 467 Zhongshan Rd, Dalian 116023, Liaoning, Peoples R China |
| 推荐引用方式 GB/T 7714 | Shi, Meng,Geng, Xuhui,Wang, Chengye,et al. Quantification of Low Copy Number Proteins in Single Cells[J]. ANALYTICAL CHEMISTRY,2019,91(18):11493-11496. |
| APA | Shi, Meng,Geng, Xuhui,Wang, Chengye,&Guan, Yafeng.(2019).Quantification of Low Copy Number Proteins in Single Cells.ANALYTICAL CHEMISTRY,91(18),11493-11496. |
| MLA | Shi, Meng,et al."Quantification of Low Copy Number Proteins in Single Cells".ANALYTICAL CHEMISTRY 91.18(2019):11493-11496. |
入库方式： OAI收割
来源：大连化学物理研究所
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