Super-sensitive time-resolved fluoroimmunoassay for thyroid-stimulating hormone utilizing europiumIII nanoparticle labels achieved by protein corona stabilization, short binding time, and serum preprocessingReport as inadecuate




Super-sensitive time-resolved fluoroimmunoassay for thyroid-stimulating hormone utilizing europiumIII nanoparticle labels achieved by protein corona stabilization, short binding time, and serum preprocessing - Download this document for free, or read online. Document in PDF available to download.

Analytical and Bioanalytical Chemistry

, Volume 409, Issue 13, pp 3407–3416

First Online: 16 March 2017Received: 28 August 2016Revised: 17 February 2017Accepted: 27 February 2017DOI: 10.1007-s00216-017-0284-z

Cite this article as: Näreoja, T., Rosenholm, J.M., Lamminmäki, U. et al. Anal Bioanal Chem 2017 409: 3407. doi:10.1007-s00216-017-0284-z

Abstract

Thyrotropin or thyroid-stimulating hormone TSH is used as a marker for thyroid function. More precise and more sensitive immunoassays are needed to facilitate continuous monitoring of thyroid dysfunctions and to assess the efficacy of the selected therapy and dosage of medication. Moreover, most thyroid diseases are autoimmune diseases making TSH assays very prone to immunoassay interferences due to autoantibodies in the sample matrix. We have developed a super-sensitive TSH immunoassay utilizing nanoparticle labels with a detection limit of 60 nU L in preprocessed serum samples by reducing nonspecific binding. The developed preprocessing step by affinity purification removed interfering compounds and improved the recovery of spiked TSH from serum. The sensitivity enhancement was achieved by stabilization of the protein corona of the nanoparticle bioconjugates and a spot-coated configuration of the active solid-phase that reduced sedimentation of the nanoparticle bioconjugates and their contact time with antibody-coated solid phase, thus making use of the higher association rate of specific binding due to high avidity nanoparticle bioconjugates.

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AbstractWe were able to decrease the lowest limit of detection and increase sensitivity of TSH immunoassay using EuIII-nanoparticles. The improvement was achieved by decreasing binding time of nanoparticle bioconjugates by small capture area and fast circular rotation. Also, we applied a step to stabilize protein corona of the nanoparticles and a serum-preprocessing step with a structurally related antibody.

KeywordsSandwich-type immunoassay Time-resolved fluoroimmunoassay Nanoparticle bioconjugate Immunoassay interference Nanoparticle protein corona AbbreviationsHABHuman autoantibody

HAMAHuman anti-mouse antibody

LLDLowest limit of detection

TSHThyroid-stimulating hormone

Electronic supplementary materialThe online version of this article doi:10.1007-s00216-017-0284-z contains supplementary material, which is available to authorized users.





Author: Tuomas Näreoja - Jessica M. Rosenholm - Urpo Lamminmäki - Pekka E. Hänninen

Source: https://link.springer.com/



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