Use of biotinylated 17β-estradiol in enzyme-immunoassay development: Spacer length and chemical structure of the bridge are the main determinants in simultaneous streptavidin–antibody binding

Lacorn, Markus; Fleischer, Kerstin; Willig, Stephanie; Gremmel, Sabine; Steinhart, Hans; Claus, R.

doi:10.1016/j.jim.2004.12.011

Cited by 13 publications

(7 citation statements)

References 33 publications

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“…The biotinylation of proteins and nucleic acids is a mature technology, yet relatively few applications employing biotinylated analogues of small bioactive molecules to identify protein targets have been described. Several biotinylated steroid hormones conjugates have been generated, primarily as immunoassay probes – , with relatively few examples using these reagents to study binding with cellular receptors – . The potential value of this approach is evident in recent reports employing biotinylated analogues of small molecules to identify penicillin-binding proteins and targets of anti-inflammatory drugs, and investigate the actions of cancer drugs targeting DNA methyltransferases – .…”

Section: Discussionmentioning

confidence: 99%

Bisphenol A Directly Targets Tubulin to Disrupt Spindle Organization in Embryonic and Somatic Cells

et al. 2008

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There is increasing concern that animal and human reproduction may be adversely affected by exposure to xenoestrogens that activate estrogen receptors. There is evidence that one such compound, Bisphenol A (BPA), also induces meiotic and mitotic aneuploidy, suggesting that these kinds of molecules may also have effects on cell division. In an effort to understand how Bisphenol A might disrupt cell division, a phenotypic analysis was carried out using sea urchin eggs, whose early embryonic divisions are independent of zygotic transcription. Fertilized Lytechinus pictus eggs exposed to BPA formed multipolar spindles resulting in failed cytokinesis in a dose-dependent, transcriptionally independent manner. By use of novel biotinylated BPA affinity probes to fractionate cell-free extracts, tubulin was identified as a candidate binding protein by mass spectrometry, and BPA promoted microtubule polymerization and centrosome-based microtubule nucleation in vitro but did not appear to display microtubule-stabilizing activity. Treatment of mammalian cells demonstrated that BPA as well as a series of Bisphenol A derivatives induced ectopic spindle pole formation in the absence of centrosome overduplication. Together, these results suggest a novel mechanism by which Bisphenol A affects the nucleation of microtubules, disrupting the tight spatial control associated with normal chromosome segregation, resulting in aneuploidy.

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Section: Discussionmentioning

confidence: 99%

Bisphenol A Directly Targets Tubulin to Disrupt Spindle Organization in Embryonic and Somatic Cells

et al. 2008

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“…The chemical structure of the linker as well as the coupling position will very much affect the functionality of the hapten molecule. The introduction of the a-amino group at position C6 of the estradiol molecule of the panning antigen has been suggested to change the hapten structure, 30 resulting in the selected scFv being non-responsive to the soluble unmodified hapten. Thus, the inability to find clones recognizing soluble hapten in this case is more likely an inherent property of the hapten itself and not of the library per se.…”

Section: Discussionmentioning

confidence: 99%

A Focused Antibody Library for Improved Hapten Recognition

Persson

Lantto

Ohlin

2006

Journal of Molecular Biology

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“…The antibody coupled to the solid phase showed effective affinity reaction to the antigen and the conjugate. Several conjugates have been previously described for antigen and biotin acting as a bridge for EIA (Lacorn, Fleischer, Wiling, Steinhart, & Claus, 2005;Mares, DeBoever, Stans, Bosmans, & Kohen, 1995). These conjugates are of defined chemical structure, but, since there is only one residue for biotinylation, there is no amplification effect as the one described here.…”

Section: Eia Proceduresmentioning

confidence: 95%

Development of an antigen-biotin conjugate with a protein bridge and its application for the development of an enzyme immunoassay

Spotorno

Tezón

2012

Food and Agricultural Immunology

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Antigens with aromatic amino groups can be conjugated to proteins by a diazo bond. This property was used to synthesise an antigen-biotin conjugate using bovine serum albumin (BSA) as a bridge, and tested this conjugate in an enzymoimmunoassay for clenbuterol detection. The assay included a highly specific antibody and the biotin-avidin amplification system. Its coefficient of variation was lower than 20% in the range of 100Á8000 pg per well and the limit of detection was 6.4 pg. The synthesis of clenbuterolÁBSAÁbiotin conjugate was reproducible, the reaction product easily purified and the amplification capacity could be set by adjusting the clenbuterol:biotin ratio.

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Use of biotinylated 17β-estradiol in enzyme-immunoassay development: Spacer length and chemical structure of the bridge are the main determinants in simultaneous streptavidin–antibody binding

Cited by 13 publications

References 33 publications

Bisphenol A Directly Targets Tubulin to Disrupt Spindle Organization in Embryonic and Somatic Cells

Bisphenol A Directly Targets Tubulin to Disrupt Spindle Organization in Embryonic and Somatic Cells

A Focused Antibody Library for Improved Hapten Recognition

Development of an antigen-biotin conjugate with a protein bridge and its application for the development of an enzyme immunoassay

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