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Synthesis and application of firefly luciferase antibody conjugates in a bioluminescent immunoassay of Salmonella cells

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Moscow University Chemistry Bulletin Aims and scope

Abstract

A method has been developed for synthesizing stable and highly active conjugates of firefly luciferase with antibodies using the heterobifunctional coupling agent N-succinimidyl 3-(2-pyridyldithio) propionate linking NH2 groups of antibodies with free SH groups of thermostable Luciola mingrelica firefly luciferase (Luc). A novel specific matrix for sorption of Salmonella cells has been designed that includes monodisperse polystyrene microparticles (φ = 240 nm) coated with Pluronic F108-PDS, which are then covalently linked to monoclonal antibodies against Salmonella cells (Sal). The Luc-Sal conjugates are used as detecting agents for bioluminescent immunoassay of Salmonella cells. The use of the novel matrix and highly active conjugates allow for a ∼100-fold increase in the method’s sensitivity compared with the standard sandwich assay.

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Authors and Affiliations

  1. Department of Chemistry, Moscow State University, Moscow, Russia

    G. Yu. Lomakina, A. Istrate & N. N. Ugarova

  2. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Pushchino Branch), Russian Academy of Sciences, Pushchino, Russia

    N. V. Rudenko

Authors
  1. G. Yu. Lomakina
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  2. A. Istrate
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  3. N. V. Rudenko
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  4. N. N. Ugarova
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Corresponding author

Correspondence to N. N. Ugarova.

Additional information

Original Russian Text © G.Yu. Lomakina, A. Istrate, N.V. Rudenko, N.N. Ugarova, 2014, published in Vestnik Moskovskogo Universiteta. Khimiya, 2014, No. 2, pp. 71–78.

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Lomakina, G.Y., Istrate, A., Rudenko, N.V. et al. Synthesis and application of firefly luciferase antibody conjugates in a bioluminescent immunoassay of Salmonella cells. Moscow Univ. Chem. Bull. 69, 49–55 (2014). https://doi.org/10.3103/S0027131414020047

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  • DOI: https://doi.org/10.3103/S0027131414020047

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