Bioluminescent detection probe for tick-borne encephalitis virus immunoassay

Описание

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, Immunoassay, Luciferase, Single-chain antibody, Tick-borne encephalitis virus, Antibodies, Bioluminescence, Chains, Glycoproteins, Immunology, Plants (botany), Probes, Proteins, Viruses, Biological properties, Conventional methods, Immunoassay, Luciferase, 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, immunoassay, 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, Immunoassay, Luciferases, Renilla, Luminescent Agents, Luminescent Measurements, Mice, Recombinant Fusion Proteins, Single-Chain Antibodies, 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 fraПоказать полностьюgment (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.

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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

Авторы

  • 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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