Synthesis and application of firefly luciferase antibody conjugates in a bioluminescent immunoassay of Salmonella cells
Autor: | G. Yu. Lomakina, N. V. Rudenko, Alena Istrate, N.N. Ugarova |
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Rok vydání: | 2014 |
Předmět: | |
Zdroj: | Moscow University Chemistry Bulletin. 69:49-55 |
ISSN: | 1935-0260 0027-1314 |
Popis: | 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. |
Databáze: | OpenAIRE |
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