Development of an Open Sandwich ELISA for the Detection of Microcystin-LR
| Autor: | Jinhua Dong, Chen Limei, Yongmei Zhou, Xinyu Li, Haimei Li, Panpan Sun, Ruyang Tan, Liqian Zhang, Hiroshi Ueda, Shengshuo Zhang, Yang Cong |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Detection limit
chemistry.chemical_classification Phage display biology Chemistry Toxin 010401 analytical chemistry Microcystin-LR Maltose binding 02 engineering and technology Microcystin 021001 nanoscience & nanotechnology Immunoglobulin light chain medicine.disease_cause 01 natural sciences Molecular biology 0104 chemical sciences Analytical Chemistry chemistry.chemical_compound medicine biology.protein Antibody 0210 nano-technology Spectroscopy |
| Zdroj: | Microchemical Journal. 158:105325 |
| Popis: | Microcystin with leucine and arginine (MCLR) is a hepatotoxic toxin produced by cyanobacteria present in watercourses that causes cell death and tissue necrosis at high exposure levels. In this study, a sensitive non-competitive, Open sandwich Enzyme-linked Immunosorbent Assay (OS-ELISA) with wide detectable range was developed for the quantification of MCLR. OS-ELISA is based on the principle of antigen-driving interaction enhancement between variable regions of an antibody. The genes for the variable regions of antibody 3A8 against MCLR were used to construct the phage display vector pDong1/Fab(3A8) to prepare assay elements. The OS-ELISA performed with phage-displayed antibody VH fragment and secreted light chain showed a limit of detection (LOD) of 0.14 nM and a wide working range from 0.14 to 10,000 nM MCLR. Another format of OS-ELISA that used purified Maltose Binding Protein-fused VL and VH-phage showed a lower LOD of 85 pM MCLR. Using the developed assay, the recovery rates for samples of lake or river water spiked with MCLR were found to range from 95.5% to 116.4%, suggesting that the assay has good potential to be used as a monitoring tool for MCLR in nature. |
| Databáze: | OpenAIRE |
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