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

Журнал: ANALYTICAL AND BIOANALYTICAL CHEMISTRY

Выпуск журнала: Vol. 401, Is. 8

Номера страниц: 2573-2579

ISSN журнала: 16182642

Место издания: HEIDELBERG

Издатель: SPRINGER HEIDELBERG

Авторы

  • 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)

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