Quantitative analysis of cholinesterase from Daphnia magna by indirect and non-competitive enzyme-linked immunosorbent assay
Polyclonal anti-serum of cholinesterase with dilution ratio of 1∶8 000 was obtained by immunizing BALB/c mice with purified cholinesterase (ChE) from Daphnia magna. An indirect and non-competitive enzyme-linked immunosorbent assay (ELISA), which used mice anti-Daphnia ChE as antibodies to react with...
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| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2012-05-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.03.016 |
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| Summary: | Polyclonal anti-serum of cholinesterase with dilution ratio of 1∶8 000 was obtained by immunizing BALB/c mice with purified cholinesterase (ChE) from Daphnia magna. An indirect and non-competitive enzyme-linked immunosorbent assay (ELISA), which used mice anti-Daphnia ChE as antibodies to react with the Daphnia ChE, was established to quantify immunoreactive content of ChE in D. magna. The minimal detection limit of the method was estimated to be 24 ng·mL<sup>-1</sup>. Specificity of anti-serum was high, with cross reaction of <0.25%, <0.25%, 2.13%, 10.19%, 3.88%, 3.56%, 2.81%, 3.93%, 5.00% and 2.75% towards non-ChE marker (5, 10 μg·mL<sup>-1</sup>), Chironomus kiinensis, A lona sp., Brachydanio rerio (the brain, the whole body without brain), Bombyx mori, A pis mellifera L., Eisenia foetida, and tadpoles of Xenopus laevis, respectively. It is indicated that the established method can be used to quantify immunoreactive content of ChE in D. magna so as to measureme accurately in vivo activity of the ChE in pesticide exposure. |
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| ISSN: | 1008-9209 2097-5155 |