Development of zeptomole and attomolar detection sensitivity of biotin-peptide using a dot-blot goldnanoparticle immunoassay

被引:74
作者
Hou, Shao-Yi
Chen, Hsi-Kuei
Cheng, Hsu-Chieh
Huang, Chun-Yen
机构
[1] Natl Taiwan Univ, Dept Chem Engn, Taipei 106, Taiwan
[2] Natl Taiwan Univ, Inst Biotechnol, Taipei 106, Taiwan
[3] Hwa Hsia Inst Technol, Dept Biochem Engn, Chungho 235, Taiwan
关键词
D O I
10.1021/ac061507g
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
An ultrasensitive, simple, and fast immunoassay for biotin-peptide detection using gold nanoparticles conjugated with antibodies has been developed. Biotin was covalently attached to a peptide and the biotin-peptide bound on a nitrocellulose membrane. Antibody-coated gold nanoparticles bound to the biotin-peptide formed red dots. With this method, 100 amol of the biotin-peptide was detected and no immunogold was bound to the membrane in the absence of biotin. The relative intensity of each dot was scored using Quantity One, a quantitative analysis software program. The linear working range of this assay was between 1 pmol and 1 mu mol. The assay sensitivity was increased by silver enhancement to 100 zmol, and the linear working range was between 100 zmol and 100 fmol. This assay can be extended to detect target molecules, such as dioxin, digoxin, mercury, and so on, with matched antibodies and has potential broad applications in immunoassay.
引用
收藏
页码:980 / 985
页数:6
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