Development of a quantitative immuno-polymerase chain reaction assay to detect and quantify low levels of human thyroid stimulating hormone

Abud, Julián Elías; Santamaría, Clarisa Guillermina; Luque, Enrique H.; Rodríguez, Horacio Adolfo

doi:10.1016/j.ab.2017.10.023

Cited by 3 publications

(1 citation statement)

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“…The quantification of circulating IFN-a2b in plasma samples was carried out by an iPCR assay based on Niemeyer et al [25]. Coating was achieved overnight, at 4 • C, in polypropylene 48-well plates, as described by Abud et al [26,27], by incubating the plates with 100 ng per well of the anti-rhIFN-α2b mAb diluted in 50 mM carbonate/bicarbonate buffer (pH 9.6). After blocking 1 h at room temperature (RT) with 1% (w/v) BSA in PBS, plates were incubated with twofold serial dilutions of rhIFN-α2b from 2000 to 15.625 pg/ml (standard curve) or the plasma sample dilutions, during 1 h at RT.…”

Section: Immuno-pcr Assaymentioning

confidence: 99%

Design and Validation of an Immuno-PCR Assay for IFN-Α2B Quantification in Human Plasma

et al. 2019

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Aim: Nowadays, IFN-α is considered a promising therapeutic target for systemic lupus erythematosus. An immuno-PCR (iPCR) was developed to quantify low amounts of IFN-α in human plasma followed by a deep analysis of the methodologic robustness throughout quality by design approach. Results: An accurate, sensitive, selective and versatile iPCR was validated. The critical iPCR procedural steps were identified, applying a Plackett–Burman design. Also, this assay demonstrated an outstanding LOD of 0.3 pg/ml. A significant aspect relies on its high versatility to detect and quantify other cytokines in human plasma as the appropriate biotinylated antibody is employed. Conclusion: This reliable iPCR assay can be clinically used as an alternative method for quantitating and detecting low IFN-α2b concentrations in human plasma samples.

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Section: Immuno-pcr Assaymentioning

confidence: 99%