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