Interference in free thyroxine immunoassay

Ghosh, Sujoy; Howlett, M. R.; Boag, D E; Malik, Iqbal; Collier, Andrew

doi:10.1016/j.ejim.2007.05.009

Cited by 21 publications

(10 citation statements)

References 6 publications

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“…It is possible that the RIA methodology may be overestimating circulating thyroid hormone levels due to interferences with antibodies or other plasma matrices as has been previously reported with thyroid hormone-immunoassay based approaches in clinical settings [20, 22, 55]. For instance, heterophilic antibodies in plasma or serum can promote non-specific binding to anti-T4 and anti-T3 antibodies (and other thyroid-targeted antibodies) producing abnormal values in RIAs.…”

Section: Resultsmentioning

confidence: 99%

“…While sensitive, immunoassay approaches have several weaknesses, including that they can be compromised by a lack of specificity and accuracy due to analytical interferences. Biomolecules may vary across species, cross-react with antibodies, and interfere with assay performance [20–22]. Moreover, RIA methods measure thyroid hormones individually thereby limiting thorough evaluations of interactive thyroid hormone homeostasis.…”

Section: Introductionmentioning

confidence: 99%

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Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS

et al. 2013

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Thyroid hormones are critical regulators of normal development and physiological functioning in all vertebrates. Radioimmunoassay (RIA) approaches have been the method of choice for measuring circulating levels of thyroid hormones in vertebrates. While sensitive, RIA-based approaches only allow for a single analyte measurement per assay, can lack concordance across platforms and laboratories, and can be prone to analytical interferences especially when used with fish plasma. Ongoing advances in liquid chromatography tandem mass spectrometry (LC/MS/MS) have led to substantial decreases in detection limits for thyroid hormones and other biomolecules in complex matrices, including human plasma. Despite these advances, current analytical approaches do not allow for the measurement of native thyroid hormone in teleost fish plasma by mass spectrometry and continue to rely on immunoassay. In this study, we developed a new method that allows for the rapid extraction and simultaneous measurement of total T4 (TT4) and total T3 (TT3) in low volumes (50 μL) of fish plasma by LC/MS/MS. Methods were optimized initially in plasma from rainbow trout (Oncorhynchus mykiss) and applied to plasma from other teleost fishes, including fathead minnows (Pimephales promelas), mummichogs (Fundulus heteroclitus), sockeye salmon (Oncorhynchus nerka), and coho salmon (Oncorhynchus kisutch). Validation of method performance with T4- and T3-spiked rainbow trout plasma at 2 and 4 ng/mL produced mean recoveries ranging from 82 to 95 % and 97 to 105 %, respectively. Recovery of 13C12-T4 internal standard in plasma extractions was: 99±1.8 % in rainbow trout, 85±11 % in fathead minnow, 73±5.0 % in mummichog, 73±1.7 % in sockeye salmon, and 80±8.4 % in coho salmon. While absolute levels of thyroid hormones measured in identical plasma samples by LC/MS/MS and RIA varied depending on the assay used, T4/T3 ratios were generally consistent across both techniques. Less variability was measured among samples subjected to LC/MS/MS suggesting a more precise estimate of thyroid hormone homeostasis in the species targeted. Overall, a sensitive and reproducible method was established that takes advantage of LC/MS/MS techniques to rapidly measure TT4 and TT3 with negligible interferences in low volumes of plasma across a variety of teleost fishes.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS

et al. 2013

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“…Such antibodies commonly affect sandwich ELISAs by cross‐linking the assay capture and detection antibodies, leading to increased signals. In the worst‐case scenario, the outcome is the misinterpretation of test results and misdiagnosis, as has been reported in human medicine …”

Section: Introductionmentioning

confidence: 94%

Prevalence of interfering antibodies in dogs and cats evaluated using a species‐independent assay

Bergman

Larsson

Hansson-Hamlin

et al. 2018

Veterinary Clinical Pathol

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“…However, due to differences in reagent specificity the concentration of free thyroxine in a given specimen determined with assays from different manufacturers can vary. In addition, heterophile antibody interference with thyroxine quantification that caused clinical confusions has been reported (2,3). To improve the reliability of assay results, mass spectrometry –based analytical protocols have been recently developed (4,5).…”

Section: Introductionmentioning

confidence: 99%

A Homogeneous Immunoassay of Thyroxine Based on Microchip Electrophoresis and Chemiluminescence Detection

Zhao

Liu

2012

Methods in Molecular Biology

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Summary A homogenous chemiluminescent immunoassay of thyroxine (T4) present in serum samples is described. The proposed method deployed the competitive immunoreaction of T4 and horseradish peroxidase (HRP)-labeled T4 (HRP–T4) with anti-T4 mouse monoclonal antibody (Ab). HRP–T4 and the HRP–T4–Ab complex were separated and quantified by using microchip electrophoresis (MCE) with chemiluminescence (CL) detection. The MCE separation was accomplished within 60 sec. Highly sensitive CL detection was achieved by means of HPR-catalyzed luminol-H2O2 reaction. The linear range for T4 were 5 −250 nM with a detection limit of 2.2 nM (S/N = 3).

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Interference in free thyroxine immunoassay

Cited by 21 publications

References 6 publications

Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS

Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS

Prevalence of interfering antibodies in dogs and cats evaluated using a species‐independent assay

A Homogeneous Immunoassay of Thyroxine Based on Microchip Electrophoresis and Chemiluminescence Detection

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