Studies on the Influence of the Structural Modifications in the Tracer on the Immunoassay of Progesterone

Karir, T.; Samuel, Grace; Kothari, Kanchan; Sivaprasad, N.; Venkatesh, Meera

doi:10.1080/15321810600573085

Cited by 12 publications

(2 citation statements)

References 3 publications

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“…With the heterologous tracer 5-OH-DCF-AMF, approximately the same sensitivity as with the homologous tracer DCF-Ahx-AMF was obtained (compare Table 2 ). According to some studies [ 27 – 29 ], heterologous tracers may improve the sensitivity. Yet, many tracers built on heterologous structures will not bind to the antibody (see [ 30 ]).…”

Section: Resultsmentioning

confidence: 99%

Fluorescence polarization immunoassay for the determination of diclofenac in wastewater

et al. 2020

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Pharmacologically active compounds are often detected in wastewater and surface waters. The nonsteroidal anti-inflammatory drug diclofenac (DCF) was included in the European watch list of substances that requires its environmental monitoring in the member states. DCF may harmfully influence the ecosystem already at concentrations ≤ 1 μg L−1. The fast and easy quantification of DCF is becoming a subject of global importance. Fluorescence polarization immunoassay (FPIA) is a homogeneous mix-and-read method which does not require the immobilization of reagents. FPIA can be performed in one phase within 20–30 min, making it possible to analyse wastewater without any complicated pre-treatment. In this study, new tracer molecules with different structures, linking fluorophores to derivatives of the analyte, were synthesized, three homologous tracers based on DCF, two including a C6 spacer, and one heterologous tracer derived from 5-hydroxy-DCF. The tracer molecules were thoroughly assessed for performance. Regarding sensitivity of the FPIA, the lowest limit of detection reached was 2.0 μg L−1 with a working range up to 870 μg L−1. The method was validated for real wastewater samples against LC-MS/MS as reference method with good agreement of both methods.

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Section: Resultsmentioning

confidence: 99%

Fluorescence polarization immunoassay for the determination of diclofenac in wastewater

et al. 2020

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“…[18] Bridge heterology in steroid immunoassay improves the assay sensitivity if 125 I, enzymes, and chemiluminescent compounds are used as labels. [19][20][21][22] The effect of linkers in enzyme conjugates has been studied so far with respect to their increasing length. However, there is a relative dearth of literature discussing the effect of varying the length of linkers in the label on the sensitivity, ED 50 , and specificity of DHEA-3-HS-BSA antibody.…”

Section: Introductionmentioning

confidence: 99%

Influence of Different Length Linker Containing Dhea-7-Cmo-Enzyme Conjugates on Sensitivity and Specificity of Dhea-3-Hs-Antibody

Shrivastav

Chaube

Kariya³

et al. 2012

Journal of Immunoassay and Immunochemistry

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The introduction of spacers in coating steroid antigen or enzyme conjugates or immunogen is known to exert an influence on the sensitivity of steroid enzyme immunoassays. We have introduced different homobifunctional spacers having varying atomic length (3 to 10) between enzyme and dehydroepiandrosterone (DHEA) moiety and studied their effects on functional parameters such as sensitivity and specificity of DHEA enzyme immunoassays. DHEA-3-hemisuccinate-bovine serum albumin (DHEA-3-HS-BSA) was used as immunogen to raise the antiserum in New Zealand white rabbits. Five enzyme conjugates were prepared using DHEA-7-carboxymethyloxime (DHEA-7-CMO) as carboxylic derivative of DHEA and horseradish peroxidase (HRP) as an enzyme label. These were DHEA-7-CMO-HRP, DHEA-7-CMO-urea-HRP (DHEA-7-CMO-U-HRP), DHEA-7-CMO-ehylenediamine-HRP (DHEA-7-CMO-EDA-HRP), DHEA-7-CMO-carbohydrazide-HRP (DHEA-7-CMO-CH-HRP), and DHEA-7-CMO-adipic acid dihydrazide-HRP (DHEA-7-CMO-ADH-HRP). The influence of different atomic length linkers on sensitivity and specificity were studied with reference to label without linker. The results of the present investigation revealed that DHEA moiety having a 3-hemisuccinate carboxyl arm that is hydrophilic in nature and spacer arm urea that is also hydrophilic in nature when used for the link to the protein carrier and enzyme for the preparation of immunogen and enzyme conjugate respectively resulted in development of assay having comparable sensitivity and lowest ED(50) as compared to other spacers. Thus sensitivity and ED(50) of the assay depend partly on the nature of the steroid and spacer arm link to the carrier protein and the enzyme.

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Comparison study between direct and indirect labeling of estradiol for radioimmunoassay purpose

Sallam

Mehany

2009

J Radioanal Nucl Chem

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Studies on the Influence of the Structural Modifications in the Tracer on the Immunoassay of Progesterone

Cited by 12 publications

References 3 publications

Fluorescence polarization immunoassay for the determination of diclofenac in wastewater

Fluorescence polarization immunoassay for the determination of diclofenac in wastewater

Influence of Different Length Linker Containing Dhea-7-Cmo-Enzyme Conjugates on Sensitivity and Specificity of Dhea-3-Hs-Antibody

Comparison study between direct and indirect labeling of estradiol for radioimmunoassay purpose

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