An enzyme-linked immunosorbent assay (ELISA) for the measurement of serum antibodies to growth hormone

Wilkin, Terence J.; Casey, Cecily; Tuck, Angela; Englefield, P.

doi:10.1016/0022-1759(89)90342-6

Cited by 5 publications

(7 citation statements)

References 10 publications

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“…For all tests, 30 mL of purified IgGs, diluted ten times in a running buffer, were applied at a flow rate of 20 mL min 21 with HBS-EP as the running buffer. Regeneration was performed with 30 mL of a solution containing 5 mM NaOH and 200 mM NaCl at a flow rate of 20 mL min 21 . Analyses were performed at 25 uC.…”

Section: Biosensor Immunoassay (Bia)mentioning

confidence: 99%

“…2,8,9 Various methods have been developed and are used routinely in official racing laboratories to screen for GH abuse, such as quantification of secondary markers of GH administration: insulin-like growth factor 1 (IGF-1) [10][11][12] or IGF binding protein 3 (IGFBP-3). 13 The effects of exercise, training, circadian rhythm, age and sex on IGF-1 in the horse are now well documented 11,12 and enable IGF-1 detection above a threshold currently proposed at 860 ng mL 21 .…”

Section: Introductionmentioning

confidence: 99%

“…[14][15][16] Antibodies raised against animal growth hormones have also been reported in patients treated with human growth hormone 17,18 and slight modifications on growth rate have been shown. 19,20 In this context, three tests have been described in the literature to detect the presence of antibodies in patients treated with hGH: one method widely used for 25 years is based on chromatoelectrophoresis using the GH-I 131 tracer, 15 while enzyme-linked immunosorbent assay (ELISA) 21,22 and a High-Performance protein G Affinity Chromatography (HPAC) method using fluorescence detection 23 have also been used. In breeding animals, production of growth hormone antibodies upon recombinant GH administration is well documented in cows 24,25 and is detected by Western Blot analysis 28 days after recombinant bovine growth hormone (rbGH) administration.…”

Section: Introductionmentioning

confidence: 99%

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Detection of secondary biomarker of met-eGH as a strategy to screen for somatotropin misuse in horseracing

Bailly-Chouriberry

Chu‐Van²,

Pinel

et al. 2008

Analyst

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Since the Australian commercialisation of the recombinant equine growth hormone (reGH) in 1998 (EquiGen-5), Bresagen), this reGH, which differs only from eGH by an additional methionine at the N-terminal end (met-eGH), is worldwide suspected to be administered to racehorses as a doping agent. Indeed, the use of this biological drug is considered as a threat to horseracing since it acts both on growth, development or reproductive functions, and on the improvement of performances. In this work, we describe two reliable techniques based on surface plasmon resonance biosensor immunoassay (SPR-BIA) and solid-phase enzyme-linked immunosorbent assay (ELISA) as new, rapid and efficient long-term screening methods applicable to horseracing antidoping analysis. The ELISA and SPR-BIA tests were applied to octanoic acid purified IgGs from serum/plasma samples collected on two thoroughbreds treated with recombinant equine growth hormone for a period of two weeks. The first kinetic study of serum/plasma antibodies raised as a consequence of recombinant equine growth hormone administrations, which allows the detection from eight days up to 200 days after the beginning of the treatment, was performed. In order to trace the occurrence of anti-reGH antibodies in routine analysis and to monitor the animal level exposure to this forbidden molecule, a random population study was conducted on 233 post-race horses.

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Section: Biosensor Immunoassay (Bia)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Detection of secondary biomarker of met-eGH as a strategy to screen for somatotropin misuse in horseracing

Bailly-Chouriberry

Chu‐Van²,

Pinel

et al. 2008

Analyst

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“…Coating buffer for the ELISA was 0.1 mol/l carbonate-bicarbonate buffer, pH 9.6. Substrate buffer consisted of citrate-phosphate buffer made up as previously described [13]. The substrate for the enzyme reac tion was o-phenylenediamine (OPD).…”

Section: Methodsmentioning

confidence: 99%

“…In another set of tests, the ELISA was carried out in the conven tional manner [IO,13] with Tg added to the assay diluent and the di luted sera incubated with the Tg overnight at 4°C.…”

Section: Tg-sepharose 4b Immunoadsorbentmentioning

confidence: 99%

Improved ELISA for Thyroid Microsomal Auto-Antibodies

Ohwovoriole

Wilkin

Scott-Morgan

et al. 1988

Int Arch Allergy Immunol

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Thyroid microsomal antibodies (TMA) and thyroglobulin antibodies (TGA) are strongly associated with auto-immune thyroid disease. TMA and TGA have been mostly detected by means of either immunofluorescence (IF), tanned red cell haemagglutination (TRCH), or radio-immunoassay (RIA) until the recent development of the enzyme-linked immunosorbent assay (ELISA). The ELISA has not been as extensively used in TMA detection as in the assay for TGA. The RIA method, though more sensitive, is technically and materially very demanding while the TRCH and IF are simpler to perform but are less sensitive. The main problem with the ELISA for the detection of TMA appears to be the interference of TGA present in some test sera reacting with thyroglobulin present as a contaminant in the thyroid microsomal preparation. In this study, we compared the TMA results from an ELISA system designed to eliminate TGA interference with those of IF and TRCH. The ELISA system in which TGA interference was eliminated without concurrent inhibition of the test reaction that gives rise to false negatives was more sensitive than either IF or TRCH. A significant number of samples was falsely reported as negative by both TRCH and IF. The correlation of the degree of positivity between TRCH and ELISA was moderate and was higher than that between ELISA and IF, though both were highly significant. The ELISA technique for TMA detection described here is a more efficient, more sensitive and also more cost-effective system than either TRCH or IF, both of which should now be replaced by ELISA, provided steps are taken to avoid false positives and false negatives.

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Immunolocalization of long-chain acyl-CoAs in plant cells

Diakou

Fédou

Carde

et al. 2002

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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Long chain acyl-Coenzyme A esters (acyl-CoAs) are key substrates in many enzymic reactions of lipid metabolism. Due to their amphiphilic nature, the membrane localization of these molecules cannot be established by subcellular membrane fractionation and usual biochemical studies. We have developed another approach based on ultrastructural immunogold cytochemistry. To preserve the acyl-CoA membrane content, the plant material was freeze substituted and cryoembedded after short aldehyde fixation followed by quick freezing. Using Arabidopsis thaliana root cells and specific antibodies raised against acyl-CoAs, we show that acyl-CoAs are mainly localized in endoplasmic reticulum membranes. Our results demonstrate the value of cryo-methods for the accurate localization of labile metabolites in plant cells.

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An enzyme-linked immunosorbent assay (ELISA) for the measurement of serum antibodies to growth hormone

Cited by 5 publications

References 10 publications

Detection of secondary biomarker of met-eGH as a strategy to screen for somatotropin misuse in horseracing

Detection of secondary biomarker of met-eGH as a strategy to screen for somatotropin misuse in horseracing

Improved ELISA for Thyroid Microsomal Auto-Antibodies

Immunolocalization of long-chain acyl-CoAs in plant cells

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