Proximity Hybridization-Regulated Immunoassay for Cell Surface Protein and Protein-Overexpressing Cancer Cells via Electrochemiluminescence

化学 电化学发光 检出限 免疫分析 生物传感器 试剂 分子生物学 细胞 电极 细胞培养 色谱法 抗体 生物物理学 生物化学 遗传学 生物 物理化学 免疫学
作者
Xiaofei Wang,Hongfang Gao,Honglan Qi,Qiang Gao,Chengxiao Zhang
出处
期刊:Analytical Chemistry [American Chemical Society]
卷期号:90 (5): 3013-3018 被引量:69
标识
DOI:10.1021/acs.analchem.7b04359
摘要

A simple electrochemiluminescence (ECL) immunoassay based on a proximity hybridization-regulated strategy was developed for highly sensitive and specific detection of cell surface protein and protein-overexpressing cancer cells. A biosensor was fabricated by self-assembling a thiolated capture ss-DNA3 (partially hybridize with ss-DNA1 and ss-DNA2) and blocking with 6-mercapto-1-hexanol on a gold electrode surface. Target protein was simultaneously bound by two ss-DNA-tagged antibody probes (DNA1-Ab1 and DNA2-Ab2), while DNA1 and DNA2 were brought in sufficient proximity and hybridized with capture DNA3 on the surface of the biosensor. After ECL signal reagent Ru(phen)32+ was intercalated into the hybridized ds-DNAs, ECL measurement was performed in the coreactant solution. A “signal on” proximity hybridization-regulated ECL immunoassay for alpha-fetoprotein (AFP) was developed. The ECL intensity increased with the increase of AFP concentration in the range of 0.05–20.0 ng/mL with a detection limit of 6.2 pg/mL. Moreover, the developed ECL method was successfully used to detect AFP-overexpressing cancer cells (MCF-7 cancer cells as model) with a detection limit of 620 cells/mL (∼60 MCF-7 cells in 100 μL of cell suspension) and discriminate AFP expression on different types of the living cell surface. This work for the first time reports a proximity hybridization-regulated ECL immunoassay for the detection of the cell surface protein on a living cell surface with good specificity and sensitivity. This simple, specific, and sensitive strategy is greatly promising for the detection of proteins and specific cells.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
chuyue完成签到,获得积分10
1秒前
正直的盼夏完成签到,获得积分10
2秒前
3秒前
4秒前
4秒前
4秒前
本特利完成签到,获得积分10
5秒前
6秒前
6秒前
6秒前
stories发布了新的文献求助20
6秒前
1234发布了新的文献求助10
7秒前
7秒前
南巷完成签到,获得积分10
8秒前
梦游游游完成签到,获得积分10
9秒前
曹乌发布了新的文献求助30
9秒前
科研通AI2S应助土豪的黑夜采纳,获得10
10秒前
隐形曼青应助morena采纳,获得10
10秒前
健忘天问发布了新的文献求助10
11秒前
11秒前
Liao发布了新的文献求助10
14秒前
落后忆丹发布了新的文献求助50
15秒前
土豪的黑夜完成签到,获得积分10
17秒前
Anri发布了新的文献求助10
17秒前
无私航空完成签到,获得积分10
18秒前
18秒前
20秒前
Owen应助健忘天问采纳,获得10
20秒前
科研小白完成签到 ,获得积分10
23秒前
29秒前
30秒前
CC完成签到,获得积分10
31秒前
32秒前
33秒前
34秒前
34秒前
34秒前
孙小雨发布了新的文献求助10
35秒前
36秒前
ll发布了新的文献求助10
36秒前
高分求助中
Evolution 10000
Sustainability in Tides Chemistry 2800
The Young builders of New china : the visit of the delegation of the WFDY to the Chinese People's Republic 1000
юрские динозавры восточного забайкалья 800
English Wealden Fossils 700
Foreign Policy of the French Second Empire: A Bibliography 500
Chen Hansheng: China’s Last Romantic Revolutionary 500
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3146297
求助须知:如何正确求助?哪些是违规求助? 2797687
关于积分的说明 7825144
捐赠科研通 2454059
什么是DOI,文献DOI怎么找? 1305990
科研通“疑难数据库(出版商)”最低求助积分说明 627630
版权声明 601503