Citation: TIAN Hui-Fang, XU Qing-Chun, DING Hui-Rong, TIAN Zhi-Hua, ZHANG Hong, ZHU-Hua, WANG Xin, SHEN Jing. New Peptide Labeling Technique Based on Bifunctional Reagent and Rare Earth Metal Chelating Tags[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(7): 1022-1028. doi: 10.11895/j.issn.0253-3820.150615 shu

New Peptide Labeling Technique Based on Bifunctional Reagent and Rare Earth Metal Chelating Tags

1.
Key Laboratory of Carcinogenesis and Translational Research of Ministry of Education/Beijing, Central Laboratory, Peking University Cancer Hospital & Institute, Beijing 100142, China;
2.
Beijing Zhonghaikang Health Technology Development Co., Ltd, Beijing 100082, China;
3.
Department of Nuclear Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China;
4.
Department of Biochemical Protection, Institute of Chemical Defense, Beijing 102205, China

Received Date: 3 October 2015
Revised Date: 11 March 2016

Figures(5)

A novel labeling approach with good compatibility based on 2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (p-SCN-Bn-DOTA) was established. The labeling conditions of p-SCN-Bn-DOTA with standard peptides were optimized, and the labeling efficiency of macrocyclic complex of p-SCN-Bn-DOTA with lanthanide metal ion Eu(Ⅲ) was also investigated. The labeling process was a two-step reaction. The labeling efficiency was affected by several factors including pH, buffer, reaction time, reaction temperature, and ratio of p-SCN-Bn-DOTA to peptide. The optimized conditions were as followed:the first step of labeling reaction was reacted in 0.2 mol/L triethylene diamine (TEAB) (pH 8.5) buffer for 60 min at 60℃, and the molar ratio of p-SCN-Bn-DOTA with peptide was 32. The second step of rare earth metal chelating was in pH 4.0 HCl/NaAC buffer for 60 min at 60℃. This approach was successfully applied in RGD peptide labeling.
- Bifunctional chelating reagents,
- Labeling,
- Peptide,
- Protein quantification,
- Mass spectrometry
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沈阳化工大学材料科学与工程学院沈阳 110142