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Moscow University Chemistry Bulletin

, Volume 69, Issue 2, pp 49–55 | Cite as

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

  • G. Yu. Lomakina
  • A. Istrate
  • N. V. Rudenko
  • N. N. UgarovaEmail author
Article

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.

Keywords

firefly luciferase Luciola mingrelica monoclonal antibody conjugation cell detection Salmonella bioluminescent enzyme immunoassay Pluronic F108 

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Copyright information

© Allerton Press, Inc. 2014

Authors and Affiliations

  • G. Yu. Lomakina
    • 1
  • A. Istrate
    • 1
  • N. V. Rudenko
    • 2
  • N. N. Ugarova
    • 1
    Email author
  1. 1.Department of ChemistryMoscow State UniversityMoscowRussia
  2. 2.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Pushchino Branch)Russian Academy of SciencesPushchinoRussia

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