Ca2+-triggered coelenterazine-binding protein from Renilla as an enzyme-dependent label for binding assay


Тип публикации: статья из журнала

Год издания: 2011

Идентификатор DOI: 10.1007/s00216-011-5343-2

Ключевые слова: Ca2+-triggered coelenterazine-binding protein (CBP), Renilla muelleri luciferase, Bioluminescent solid-phase microassay, Ca 2+-triggered coelenterazine-binding protein (CBP), Binding assays, Detection limits, Frozen state, In-vitro assays, Microassay, Model experiments, Multianalytical systems, Renilla luciferase, Solid-phase, Calcium, Chemical modification, Oligonucleotides, Proteins, coelenterazine, imidazole derivative, luminescent agent, protein, pyrazine derivative, Renilla luciferin 2 monooxygenase, animal, article, chemical structure, enzymology, luminescence, metabolism, methodology, protein binding, Renilla, Animals, Imidazoles, Luciferases, Renilla, Luminescent Agents, Luminescent Measurements, Models, Molecular, Pyrazines, Renilla muelleri

Аннотация: The recombinant Ca2+-triggered coelenterazine-binding protein (CBP) from Renilla muelleri was investigated as a biospecifically labeled molecule for in vitro assay applications. The protein was shown to be stable in solutions in the frozen state, as well as stable under heating and to chemical modifications. Conjugates with biotin,Показать полностьюoligonucleotide, and proteins were obtained and applied as biospecific molecules in a solid-phase microassay. CBP detection was performed with intact (no modifications were made) Renilla luciferase in the presence of calcium, and the detection limit was found to be 75 amol. Model experiments indicate that this approach shows much promise, especially with regard to the development of multianalytical systems.

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Выпуск журнала: Vol. 401, Is. 8

Номера страниц: 2573-2579

ISSN журнала: 16182642

Место издания: HEIDELBERG



  • Krasitskaya V.V. (Institute of Biophysics,Russian Academy of Sciences SB)
  • Markova S.V. (Institute of Biophysics,Russian Academy of Sciences SB)
  • Stepanyuk G.A. (Institute of Biophysics,Russian Academy of Sciences SB)
  • Frank L.A. (Siberian Federal University)
  • Korneeva S.I. (Siberian Federal University)
  • Kudryavtsev A.N. (Siberian Federal University)