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Biotinylation of quantum dots for application in fluoroimmunoassays with biotin-avidin amplification

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Abstract

A competitive microplate fluoroimmunoassay was developed for the determination of human serum albumin in urine. It is based on the use of biotinylated CdTe quantum dots (QDs) whose synthesis is optimised in terms of storage stability, purification, and signal-to-noise ratio. The bioconjugated QDs were characterised by gel chromatography and gel electrophoresis. Storage stability and quantum yield were investigated. The excitation/emission wavelengths are 360/620 nm. The immunoassay of human serum albumin in urine has a working range from 1.7 to 10 μg.mL−1, and the limit of detection is 1.0 μg.mL−1.

Preparation of biotinylated quantum dots is described. Their structure consists of biotinylated denatured bovine serum albumin attached to the quantum dot surface. Fluoroimmunoassay for human serum albumin was developed utilizing thus prepared bioconjugate.

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Acknowledgement

The authors gratefully acknowledged Dipl. Ing. Alois Tichý and Dr. Karel Kouřil (Clinical Laboratory of Blansko Hospital) for providing the analysis of urine samples and their general help. The research was supported by the Ministry of Education of Czech Republic under the projects “Biomolecular Centre” (LC06030) and “Proteins in metabolism and interaction of organisms with the environment” (MSM0021622413).

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Authors and Affiliations

  1. Petr Skládal, Department of Biochemistry, Masaryk University, Kotlářská 2, 611 37, Brno, Czech Republic

    Antonin Hlavacek, Pavel Bouchal & Petr Skládal

  2. CEITEC MU, Masaryk University, Brno, Czech Republic

    Petr Skládal

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  1. Antonin Hlavacek
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  2. Pavel Bouchal
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  3. Petr Skládal
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Correspondence to Petr Skládal.

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Hlavacek, A., Bouchal, P. & Skládal, P. Biotinylation of quantum dots for application in fluoroimmunoassays with biotin-avidin amplification. Microchim Acta 176, 287–293 (2012). https://doi.org/10.1007/s00604-011-0729-6

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  • DOI: https://doi.org/10.1007/s00604-011-0729-6

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