Тип публикации: статья из журнала
Год издания: 2015
Идентификатор DOI: 10.1007/s00216-015-8710-6
Ключевые слова: Tick-borne encephalitis virus, Single-chain antibody, Luciferase, Immunoassay, Antibodies, Bioluminescence, Chains, Glycoproteins, Immunology, Plants (botany), Probes, Proteins, Viruses, Biological properties, Conventional methods, Recognition element, Single chain variable fragments, Single-chain antibodies, Solid-phase immunoassay, Antigens, hybrid protein, luminescent agent, Renilla luciferin 2 monooxygenase, single chain fragment variable antibody, animal, chemistry, genetics, human, isolation and purification, luminescence, metabolism, mouse, procedures, tick, tick borne encephalitis, Tick borne encephalitis virus, virology, Animals, Encephalitis Viruses, Tick-Borne, Encephalitis, Tick-Borne, Humans, Luciferases, Renilla, Luminescent Agents, Luminescent Measurements, Mice, Recombinant Fusion Proteins, Ticks
Аннотация: To facilitate the detection of the tick-borne encephalitis virus (TBEV), the causative agent of one of the most severe human neuroinfections, we have developed an immunoassay based on bioluminescent hybrid protein 14D5a-Rm7 as a detection probe. The protein containing Renilla luciferase as a reporter and a single-chain variable fragment (scFv) of murine immunoglobulin to TBEV as a recognition element was constructed, produced by bacterial expression, purified, and tested. Both domains were shown to reveal their specific biological properties-affinity to the target antigen and bioluminescent activity. Hybrid protein was applied as a label for solid-phase immunoassay of the antigens, associated with the tick-borne encephalitis virus (native glycoprotein E or extracts of the infected strain of lab ticks). The assay demonstrates high sensitivity (0.056 ng of glycoprotein E; 10(4)-10(5) virus particles or 0.1 pg virions) and simplicity and is competitive with conventional methods for detection of TBEV.
Издание
Журнал: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Выпуск журнала: Vol. 407, Is. 18
Номера страниц: 5417-5423
ISSN журнала: 16182642
Место издания: HEIDELBERG
Издатель: SPRINGER HEIDELBERG
Персоны
- Burakova L.P. (Institute of Biophysics, Siberian Branch, Russian Academy of Sciences)
- Yakimenko V.V. (Research Institute of Natural Foci Infections)
- Kudryavtsev A.N. (Institute of Biophysics, Siberian Branch, Russian Academy of Sciences)
- Stepanyuk G.A. (Institute of Biophysics, Siberian Branch, Russian Academy of Sciences)
- Frank L.A. (Siberian Federal University)
- Baykov I.K. (Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences)
- Morozova V.V. (Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences)
- Tikunova N.V. (Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences)
- Dubova M.A. (Siberian Federal University)
- Lyapustin V.N. (Institute of Poliomyelitis and Viral Encephalitides, Russian Academy of Sciences)
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