98966-42-8Relevant articles and documents
Phosphate-triggered ratiometric fluoroimmunoassay based on nanobody-alkaline phosphatase fusion for sensitive detection of 1-naphthol for the exposure assessment of pesticide carbaryl
Chen, Zi-Jian,Wu, Hui-Ling,Shen, Yu-Dong,Wang, Hong,Zhang, Yi-Feng,Hammock, Bruce,Li, Zhen-Feng,Luo, Lin,Lei, Hong-Tao,Xu, Zhen-Lin
, (2021/10/12)
The excessive use of carbaryl has resulted in the risk of its exposure. In this study, we isolated six nanobodies (Nbs) from a camelid phage display library against the biomarker of carbaryl, 1-naphthol (1-NAP). Owing to its characteristics of easy genetic modifications, we produced a nanobody-alkaline phosphatase (Nb-CC4-ALP) fusion protein with good stability. A dual-emission system based ratiometric fluoroimmunoassay (RFIA) for quick and highly sensitive determination of 1-NAP was developed. Silicon nanoparticles (SiNPs) was used as an internal reference and for aggregation-induced emission enhancement (AIEE) of gold nanoclusters (AuNCs), while AuNCs could be quenched by MnO2 via oxidation. In the presence of ALP, ascorbic acid phosphate (AAP) can be transformed into ascorbic acid (AA), the later can etch MnO2 to recover the fluorescence of the AuNCs. Based on optimal conditions, the proposed assay showed 220-fold sensitivity improvement in comparison with conventional monoclonal antibody-based ELISA. The recovery test of urine samples and the validation by standard HPLC-FLD demonstrated the proposed assay was an ideal tool for screening 1-NAP and provided technical support for the monitoring of carbaryl exposure.
In Situ Fluorogenic Reaction Generated via Ascorbic Acid for the Construction of Universal Sensing Platform
Fan, Yongchao,Lv, Mengmeng,Xue, Yuan,Li, Jing,Wang, Erkang
, p. 6873 - 6880 (2021/05/31)
A highly fluorescent emission reaction between terephthalic acid (PTA) and ascorbic acid (AA) via simple control of the reaction temperature was first revealed with the detailed formation mechanism and various characterizations including electron paramagnetic resonance and mass spectrometry. Based on the AA-responsive emission, the alkaline phosphatase (ALP) triggered the transformation of l-ascorbic acid 2-phosphate trisodium salt to AA was integrated with the present system for developing a sensitive, selective, and universal platform. The monitoring of the activity of ALP and the fabrication of ALP-based enzyme-linked immunoassay (ELISA) with carcinoembryonic antigen (CEA) as the model target was performed. The fluorescence intensity correlated well to the CEA concentration in the ranges of 0.25-30 ng/mL, with a detection limit of 0.08 ng/mL. Such a facile protocol based on the fluorescent reaction between PTA and AA without the assistance of catalysis of nanomaterials avoided the laborious synthesis procedure and provided a direct strategy for the early clinical diagnosis coupled with ALP-related catalysis.
Fluorescence Immunoassay Based on the Alkaline Phosphatase Triggered in Situ Fluorogenic Reaction of o-Phenylenediamine and Ascorbic Acid
Zhao, Dan,Li, Juan,Peng, Chuanyun,Zhu, Shuyun,Sun, Jian,Yang, Xiurong
, (2019/02/19)
Inspired by the special reducing capability of ascorbic acid (AA), ascorbic acid 2-phosphate (AA2P) has been extensively utilized as a substrate in current alkaline phosphatase (ALP) activity assays owing to the ALP-triggered transformation of AA2P into AA. However, such assays usually require AA-related complicated and laborious synthesis and/or signal generation procedures. Herein, we report an interesting in situ fluorogenic interaction between o-phenylenediamine (OPD) and AA, which inspires us to put forward a novel and simple AA2P/OPD-participated fluorescence turn-on ALP activity assay for the first time, and then the corresponding ALP-based fluorescence enzyme-linked immunosorbent assay (ELISA) has also been developed by means of the conventional ELISA platforms. According to the convenient and facile detection process with clear response mechanism, our fluorogenic reaction-based assay exhibits good sensitivity, selectivity, and excellent sensing performance, which ensures fluorescence ELISA to potentially be applied in clinical diagnosis by employing a well-studied biomarker of hepatocellular carcinoma, α-fetoprotein (AFP) as the model analyte. Such original ELISA via in situ formation of fluorophore from scratch gives a new sight to develop other potential immunoassay platforms in early clinical diagnosis by controlling the target antigens in the near future.