Immunoassay using hapten—enzyme conjugates

Weemen, B.K. van; Schuurs, A.H.W.M.; Raymakers, H.H.Th.; Oostermeyer, M.W.

doi:10.1016/0014-5793(72)80830-5

Cited by 102 publications

(29 citation statements)

References 10 publications

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“…Enzyme-immunoassay for estrogen using peroxidase as a marker was reported by Weemen and Shuurs (1972), and Hell et al(1976).…”

Section: Discussionmentioning

confidence: 99%

“…To avoid this hazard, attempts have been made to develop nonisotopic immunoassays, such as viro-immunoassay (Andriew et al, 1975), fluorescence polarization immunoassay (Miyai et al, 1978), and enzyme-immunoassay (Weemen and Shuurs, 1972) for measurement of steroid hormones. Preliminary studies on enzymeimmunoassay of estrogen using peroxidase as a marker enzyme have been reported (Weemen and Shuurs, 1972;Hell et al, 1976) Urinary estriol is known to be an index of feto-placental function, but there are several problems about its measurement, such as the difficulty of colleting24-hr urine samples quantitatively. Measurement of serum estriol avoids these problems, and has the added advantage that it is a faster method for detecting fetal jeopardy.…”

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confidence: 99%

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An enzyme-immunoassay for estriol.

et al. 1979

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“…Enzyme-immunoassay for estrogen using peroxidase as a marker was reported by Weemen and Shuurs (1972), and Hell et al(1976).…”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

An enzyme-immunoassay for estriol.

et al. 1979

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“…Some researchers demonstrated the strong influence of combinations of antiserum and steroid-enzyme conjugate on the characteristics of P [6] and estrogen [29,30] EIA. Our homologous combination gave a cross-reactivity similar to other EIA [15,29,30] applying the same antiserum.…”

Section: Discussionmentioning

confidence: 99%

The Development of a Sensitive Enzymeimmunoassay for the Determination of Estrone and Estradiol-17.BETA. in Bovine Blood Plasma based on the Same Homologous Combination with Antiserum and Steroid-Enzyme Conjugate

Takenouchi

Oshima

Shimada

et al. 2004

The Journal of Veterinary Medical Science

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ABSTRACT. The development of a sensitive enzymeimmunoassay (EIA) for the determination of estrone (E1) and estradiol-17β (E2β) in bovine plasma is described. The assay is a homologous double-antibody EIA with E2β 17hemisuccinate (HS) as hapten for the immunoreactive reagent. The antiserum was raised against E2β 17HS bovine serum albumin conjugate in the rabbit, and E2β 17HS-horseradish peroxidase was used as steroid-enzyme conjugate. Each estrogen EIA was distinguished only by using the each working stan dard and sample for the EIA. Bovine plasma E1 and E2β were extracted and purified before EIA. The antiserum was used at 1:1,750,000 dilutions for EIA. Estrone and E2β showed high cross-reactivity with the antiserum (E1: 350.7%, E2β:100%). The sensitivities were <0.03 pg/well for E1 and <0.12 pg/well for E2 β. Recovery rates of E1 and E2β added to bovine blood plasma were 94.5% and 93.9%, respectively. The precision for EIA of estrogens was below 9.7%. The profiles of either estrogen as determined by EIA corresponded closely well with follicle dynamics in the cow during the estrous cycles and with placental function in pregnant animals. In co nclusion, our new EIA can be applied with sufficient sensitivities, recovery and precision for the routine analysis of E1 and E2β concentrations in bovine plasma. KEY WORDS: cattle, enzymeimmunoassay, estrone, estradiol-17β.

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“…Similarly, to achieve optimum conditions for the competition the conjugated hapten should have less affinity to antibody than that of analyte. Additionally the catalytic activity of haptenenzyme conjugate is also of great importance 16 . Considerable variations in the affinity, owing to the hapten structures, have been reported between homologous and heterologous ELISA [17][18] .…”

Section: Introductionmentioning

confidence: 99%

Effect of Spacer and the Enzyme-Linked Immunosorbent Assay

et al. 2016

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The effect of spacers and the enzyme-linked immunosorbent assay (ELISA) formats on the functional parameters of assays such as lower detection limit, inhibitory concentration at 50 per cent (IC 50 ), and specificity were studied. Enzyme conjugates having hydrophobic and hydrophilic spacers were prepared using O-isopropyl methylphosphonic acid (IMPA) and horseradish peroxidase (HRP) as an enzyme label. Comparison was made with reference to enzyme conjugate without any spacer. The present investigation revealed that the presence of a hydrophilic spacer in the enzyme conjugate significantly improves the sensitivity of assays. An enhanced IC 50 value achieved was 0.01 µg mL −1 for free antigen detection by direct immunoassay using hydrophilic spacers and precoating of ELISA plates by secondary antibody. The use of a hydrophilic spacer might have helped in projecting the hapten in the aqueous phase, leading to enhanced antibody binding signal and improved sensitivity of the assay.

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Immunoassay using hapten—enzyme conjugates

Cited by 102 publications

References 10 publications

An enzyme-immunoassay for estriol.

An enzyme-immunoassay for estriol.

The Development of a Sensitive Enzymeimmunoassay for the Determination of Estrone and Estradiol-17.BETA. in Bovine Blood Plasma based on the Same Homologous Combination with Antiserum and Steroid-Enzyme Conjugate

Effect of Spacer and the Enzyme-Linked Immunosorbent Assay

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