Rapid homogeneous immunoassay of peptides based on bioluminescence resonance energy transfer from firefly luciferase

被引:10
|
作者
Yamakawa, Y [1 ]
Ueda, H [1 ]
Kitayama, A [1 ]
Nagamune, T [1 ]
机构
[1] Univ Tokyo, Grad Sch Engn, Dept Chem & Biotechnol, Bunkyo Ku, Tokyo 1138656, Japan
关键词
fusion protein; bioluminescent protein; homogeneous immunoassay; peptide epitope;
D O I
10.1263/jbb.93.537
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A sensitive homogeneous immunoassay is needed in the field of clinical diagnostics. Here we propose a rapid and potentially sensitive homogeneous immunoassay of peptide epitope based on the bioluminescence resonance energy transfer (BRET). A GST-peptide tag fused to the N-terminus of firefly luciferase, and Cy3 or Cy3.5-labeled anti-peptide antibody were prepared to detect BRET from the hybrid luciferase to the fluorolabeled antibody. By measuring the spectral change due to BRET, the amount of c-myc peptide was successfully determined in a short period.
引用
收藏
页码:537 / 542
页数:6
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