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G. Yu. Lomakina, E. N. Istrate, N. V. Rudenko, N. N. Ugarova

Synthesis and assessment of firefly luciferase–Antibody conjugates for bioluminescent enzyme immunoassay of Salmonella cells

Abstract

A method was developed for synthesis of stable and highly active firefly luciferase–antibody conjugates using heterobifunctional agent – N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) – as a linker between NH2-groups of antibodies and free SH-groups of a thermostableLuciola mingrelica firefly luciferase (Luc). A new specific matrix to capture Salmonella cells was developed: monodisperse polystyrene microparticles were coated with Pluronic F108-PDS which was then covalently labeled with monoclonal anti-Salmonella antibodies (Sal). Luc-Sal conjugates were used as detection agents in bioluminescent immunoassay of Salmonella cells. The use of this matrix and highly active Luc-Sal conjugates permitted to extend the method sensitivity by ~100 times as compared to the standard "sandwich"-type immunoassay of Salmonella.
Key words: firefly luciferase; Luciola mingrelica; monoclonal antibody to Salmonella; conjugation; bioluminescent enzyme immunoassay for Salmonella; polystyrene microparticles; Pluronic F108-PDS.
Moscow University Chemistry Bulletin.
2014, Vol. 55, No. 2, P. 71
   

Copyright (C) Chemistry Dept., Moscow State University, 2002
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