title: Highly sensitive turn-on fluorescence detection of thrombomodulin based on fluorescence resonance energy transfer.
abstract: As an integral glycoprotein on the surface of endothelial cells, thrombomodulin (TM) has very high affinity for thrombin. TM has been regarded to be a marker of endothelial damage since it can be released during endothelial cell injury. In this work, a highly sensitive fluorescence method for the quantitative detection of TM was developed. TM antibody (Ab) and bovine serum albumin (BSA) were bound on gold nanoparticles (AuNPs) to construct BSA-AuNPs-Ab nanocomposites and they were characterized by transmission electron microscope and UV-vis spectrophotometry. The fluorescence of acridine orange (AO) was quenched by the prepared gold nanocomposites based on fluorescence resonance energy transfer (FRET). In the presence of TM, the fluorescence was turned on due to the effective separation of AO from the surface of gold nanocomposites. Under optimum conditions, the enhanced fluorescence intensity displayed a linear relationship with the logarithm of the TM concentration from 0.1pgmL-1to 5ngmL-1with a low detection limit of 12fgmL-1. The release of soluble thrombomodulin (sTM) by the injured HUVEC-C cells in the presence of H2O2was investigated using the proposed method. The released sTM content in the growth medium was found to be increased with the enhancement of contact time of the cells with H2O2.
authors: Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, PR China.
authors: Liyan
authors: L
authors: Kong
authors: Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, PR China.
authors: Jiaming
authors: J
authors: Zhu
authors: Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, PR China.
authors: Wen
authors: W
authors: Wang
authors: Department of Chemistry, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, PR China.
authors: Lehe
authors: L
authors: Jin
authors: Department of Chemistry, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, PR China.
authors: Yanjiao
authors: Y
authors: Fu
authors: Department of Chemistry, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, PR China.
authors: Bohui
authors: B
authors: Duan
authors: Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, PR China; Department of Chemistry, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, PR China. Electronic address: liangtan@hunnu.edu.cn.
authors: Liang
authors: L
authors: Tan
mesh_terms: D000165:Acridine Orange / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine; D013056:Spectrophotometry, Ultraviolet; D018180:Thrombomodulin / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; C546931:THBD protein, human; D018180:Thrombomodulin; D012710:Serum Albumin, Bovine; D006046:Gold; D006861:Hydrogen Peroxide; D000165:Acridine Orange
keywords: Acridine orange; Fluorescence resonance energy transfer; Gold nanocomposites; Immunoreaction; Thrombomodulin