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: LIU Hong-cui, YANG Yan-xia, LI Shao-nan
Format: Article
Language:English
Published: Zhejiang University Press 2012-05-01
Series:浙江大学学报. 农业与生命科学版
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Online Access:https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.03.016
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author LIU Hong-cui
YANG Yan-xia
LI Shao-nan
author_facet LIU Hong-cui
YANG Yan-xia
LI Shao-nan
author_sort LIU Hong-cui
collection DOAJ
description 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 &lt;0.25%, &lt;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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spelling doaj-art-85979c8f7e984d508d801c5bd6dbef792025-08-20T03:58:18ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552012-05-013834735410.3785/j.issn.1008-9209.2012.03.01610089209Quantitative analysis of cholinesterase from Daphnia magna by indirect and non-competitive enzyme-linked immunosorbent assayLIU Hong-cuiYANG Yan-xiaLI Shao-nanPolyclonal 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 &lt;0.25%, &lt;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.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.03.016<italic>Daphnia magna</italic>indirect and non-competitive enzyme-linked immunosorbent assaycholinesterase immunoreactive content
spellingShingle LIU Hong-cui
YANG Yan-xia
LI Shao-nan
Quantitative analysis of cholinesterase from Daphnia magna by indirect and non-competitive enzyme-linked immunosorbent assay
浙江大学学报. 农业与生命科学版
<italic>Daphnia magna</italic>
indirect and non-competitive enzyme-linked immunosorbent assay
cholinesterase immunoreactive content
title Quantitative analysis of cholinesterase from Daphnia magna by indirect and non-competitive enzyme-linked immunosorbent assay
title_full Quantitative analysis of cholinesterase from Daphnia magna by indirect and non-competitive enzyme-linked immunosorbent assay
title_fullStr Quantitative analysis of cholinesterase from Daphnia magna by indirect and non-competitive enzyme-linked immunosorbent assay
title_full_unstemmed Quantitative analysis of cholinesterase from Daphnia magna by indirect and non-competitive enzyme-linked immunosorbent assay
title_short Quantitative analysis of cholinesterase from Daphnia magna by indirect and non-competitive enzyme-linked immunosorbent assay
title_sort quantitative analysis of cholinesterase from daphnia magna by indirect and non competitive enzyme linked immunosorbent assay
topic <italic>Daphnia magna</italic>
indirect and non-competitive enzyme-linked immunosorbent assay
cholinesterase immunoreactive content
url https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.03.016
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AT yangyanxia quantitativeanalysisofcholinesterasefromdaphniamagnabyindirectandnoncompetitiveenzymelinkedimmunosorbentassay
AT lishaonan quantitativeanalysisofcholinesterasefromdaphniamagnabyindirectandnoncompetitiveenzymelinkedimmunosorbentassay