Hong Yang,Shanshan Zhu,Lihua Pang,Yifei Ma,Xiaoxue Liu,Jinhua Liu
ABSTRACT Using a single emission-wavelength to accurately/sensitively quantify disease markers in complex environments still faces enormous challenges. In this study, we construct a Cu 2+ -triggered wavelength-tunable fluorescent immunoassay for real-time detection of cTnI in human serum. The developed platform is based on an “on-off-on” strategy, which involves the rapid in-situ fluorogenic reaction of dopamine (DA) and phenol analogue by Cu 2+ , strong coupling between PPi and Cu 2+ as well as the specific hydrolysis of PPi into orthophosphate by alkaline phosphatase. By using cTnI as a model antigen, the assay shows good linearity relationships between the fluorescence response and cTnI concentration. Furthermore, we used different phenol analogues (1,3-dihydroxynaphthalene, 8-hydroxyjulolidine, 1,5-dihydroxynaphthalene) as fluorescence reaction substrates to achieve multi-channel (blue, green and yellow) real-time monitoring of cTnI with detection limits of 0.17 ng/mL, 0.17 ng/mL and 0.33 ng/mL, respectively. We successfully applied the wavelength-tunable ELISA to detect cTnI levels in real human serum, and the results were compared with a commercial ELISA with satisfactory outcomes. Therefore, our proposed platform demonstrates great potential in the early diagnosis of acute myocardial infarction.
Can Chen,Guotong Ruan,Yujie Sun,Lili Wang,Chengwu Zhang,Jinhua Liu
As inorganic pyrophosphatase (PPase) gets involved in the regulation of various pathophysiological procedures, rapid and effective detection of its activity is of great value for monitoring and managing PPase-related diseases. Herein, an in situ Cu 2+ -triggered wavelength-tunable fluorescent sensor for high sensitivity monitoring of PPase activity is constructed. The proposed sensor is based on the principle of Cu 2+ -triggered polymerization of dopamine (DA) and m-phenylenediamine (MPD), the stronger coupling effect of pyrophosphate ion (PPi) and Cu 2+ , and the effective hydrolysis of PPase to PPi. With the change of substituents of MPD, the wavelength of fluorescence polymer nanoparticles (FPNPs) exhibits tunable capacity from 535 nm to 565 nm. Compared with the traditional detection method, as developed sensor shows high sensitivity to PPase with a low detection limit of 0.333 mU/mL in green channel and 0.033 mU/mL in orange channel. Additionally, the sensor realizes PPase sensing in the diluted human serum with excellent recovery in both green and orange models. Furthermore, this sensor is also applicable for inhibition assay of PPase (NaF: IC 50 of 134.7 μM) and logic gates construction. Summarily, as prepared sensor provides one better way for monitoring PPase in diagnostic as well as drug screening.
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