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| - Biological Monitoring of 2,4,5-Trichlorophenol (I): Preparation of Antibodies and Development of anImmunoassay Using Theoretical Models
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| - Antibodies against 2,4,5-trichlorophenol have been prepared after theoretical and molecularmodeling chemical studies of three potential immunizing haptens with the aim to find out theone mimicking best the target analyte. Competitive direct and indirect ELISAs have beendeveloped after screening a battery of haptenized enzyme tracers and coating antigens,respectively. The relation between the degree of heterology of the competitor and the resultingimmunoassay detectability has been investigated according to the electronic similarities ofthe competitor haptens with the target analyte taking in consideration their pKa values. Thesestudies have been performed using theoretical and molecular modeling tools to find out theirelectronic distribution at their minimum energetic levels. The results suggest that thecompetitors should have a high homology to produced assays with good detectability values.On the other hand detectability improves when lowering the hapten density of the competitors.An indirect competitive ELISA has been finally selected for further investigation. Theimmunoassay has an IC50 value of 0.6 μg L-1 and a limit of detection of 0.084 μg L-1. Theselectivity of the assay is high in relation to other chlorophenols frequently present in realsamples. In contrast, the brominated analogues may also be recognized with this assay.
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