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Peleyeju Moses G. Mgedle Nande Viljoen Elvera L. Scurrel Mike S. Ray Sekhar C. 《Research on Chemical Intermediates》2021,47(10):4213-4226
Research on Chemical Intermediates - The application of microwave energy in the synthesis of catalytic materials has been indicated to induce desirable effects that lead to improved activity.... 相似文献
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Ferraz N Carnevia D Nande G Rossotti M Miguez MN Last JA Gonzalez-Sapienza G 《Analytical and bioanalytical chemistry》2007,389(7-8):2195-2202
Vitellogenin (VTG) and choriogenin (CHO) are valuable biomarkers of endocrine-disrupting compound (EDC) exposure in fish.
Existing immunoassays are limited to a few species, which restricts their use for the analysis of local wildlife sentinels.
Using C. facetum as a relevant South American model fish, this work presents a new strategy for the preparation of antibodies to VTG and CHO,
with zero cross-reactivity with fish serum components. Recombinant fragments of Cichlasoma facetum VTG (280-mer) and CHO (223-mer) were prepared by degenerate primer RT-PCR and expression in E. coli. Polyclonal and monoclonal antibodies prepared with these antigens were used to develop rapid dotblot assays for VTG and
CHO. Both the polyclonal and monoclonal antibodies prepared with the recombinant antigens reacted against the native proteins
adsorbed on to nitrocellulose allowing the set up of sensitive dotblot assays. The VTG assay was further validated with spiked
samples and purified native VTG. Exposure experiments with several estrogenic compounds revealed the potential of C. facetum as a sensitive biomonitor that produced measurable responses at concentrations of 100 ng L−1 of 17-beta-estradiol, 100 ng L−1 of ethynylestradiol, and 6.6 μg L−1 of nonylphenol. The approach described here may be applied to other native species to produce highly specific and sensitive
rapid tests. It may be particularly advantageous for species that cannot be kept in captivity or when homogeneous purification
of the immunizing proteins is particularly challenging. In conclusion, we present a novel approach to develop a strategy for
the generation of immunoassay reagents for vitellogenin (VTG) and choriogenin (CHO), which will facilitate regional studies
on the impact of endocrine-disrupting chemicals on local wildlife. 相似文献
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