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Conjugation reactions in the preparations of quantum dot-based immunoluminescent probes for analysis of proteins by capillary electrophoresis

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Abstract

A number of biologically important molecules, such as DNA, proteins, and antibodies, are routinely conjugated with fluorescent tags for high-sensitivity analyses. Here, the application of quantum dots in the place of bright and size-tunable luminophores is studied. Several selected bioconjugation reactions via zero-length cross-linkers, long-chain linkers, and oriented methods for linking of quantum dots with proteins were tested. Anti-ovalbumin, anti-proliferating cell nuclear antigen, anti-hemagglutinin, and anti-CD3 membrane protein as model antibodies and annexin V were used as high-specificity selectors. The reaction yield and efficiency of the prepared immunoluminescent probes were tested by capillary zone electrophoresis with laser-induced fluorescence detection.

Scheme of antibody-quantum dot conjugate

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Acknowledgments

The authors wish to thank Dr. Borivoj Vojtesek from Masaryk Memorial Cancer Institute for the kind gift of a purified anti-hemagglutinin. This work was supported by the following grants: The Grant Agency of the Czech Republic number 203/08/1680 and 301/11/2055, Ministry of Education, Youth and Sports number LC06023, bilateral project CZ-ES 2009-10/6 (2008CZ0017) and institute research plan AV0Z40310501.

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Authors and Affiliations

  1. Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Veveri 97, 602 00, Brno, Czech Republic

    M. Lišková, I. Voráčová, K. Klepárník, V. Hezinová, J. Přikryl & F. Foret

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  1. M. Lišková
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  2. I. Voráčová
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  3. K. Klepárník
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  4. V. Hezinová
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Correspondence to K. Klepárník.

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Published in the special issue Analytical and Bioanalytical Luminescence with Guest Editor Petr Solich.

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Lišková, M., Voráčová, I., Klepárník, K. et al. Conjugation reactions in the preparations of quantum dot-based immunoluminescent probes for analysis of proteins by capillary electrophoresis. Anal Bioanal Chem 400, 369–379 (2011). https://doi.org/10.1007/s00216-011-4700-5

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  • DOI: https://doi.org/10.1007/s00216-011-4700-5

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