Binding of homologous and heterologous isoluminol- and enzyme-labelled progesterone conjugates to monoclonal antibodies

Boever, Jozef G. De; Kohen, F.; Bosmans, Eugène

doi:10.1016/0003-2670(94)80061-8

Cited by 7 publications

(3 citation statements)

References 23 publications

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“…Mitsuma et al [16] have demonstrated an increase in the sensitivity of progesterone EIA that uses bridge heterology Influence of Length Linker on DHEA-3-HS-Antibody 13 in the label. Boever et al [17] have demonstrated an increase in the sensitivity of progesterone assay that uses different sites for label and immunogen preparation. Similarly, Shrivastav et al [24] have demonstrated an increase in the sensitivity of DHEA assay that uses different sites for label and immunogen preparation.…”

Section: Precisionmentioning

confidence: 98%

“…[35] The incorporation of spacer arms might have resulted in minimization of the conventional bridge recognition effects, decrease in the influence of local environment, and overcoming steric constraints between the two high molecular weight proteins, i.e., antibody and label. [15][16][17] The specificities of the antibodies obtained depend partly on the nature of the steroid link to the protein carrier, in particular the position on the steroid nucleus at which the linker is attached, since this determines the orientation of the steroid in the antibody combining site. For a ligand such as a steroid, the binding site is likely to consist of a hydrophobic cavity or pocket in which most of the ligand surface becomes buried.…”

Section: Precisionmentioning

confidence: 99%

“…[13,14] In the antibody-immobilized format, linkers have been coupled between the steroid derivative and label protein to minimize the conventional bridge recognition effects and to overcome steric constraints between the two high molecular weight proteins, i.e., antibody and label. [15][16][17] A mass-action model has been proposed to theoretically predict such behavior of assays showing differential affinities. [18] Bridge heterology in steroid immunoassay improves the assay sensitivity if 125 I, enzymes, and chemiluminescent compounds are used as labels.…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Precisionmentioning

confidence: 98%