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The Ca2+-Regulated Photoprotein Obelin as a Tool for SELEX Monitoring and DNA Aptamer Affinity Evaluation |
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| Authors: | Vasilisa V Krasitskaya Natalia S Goncharova Vladislav V Biriukov Eugenia E Bashmakova Marsel R Kabilov Ivan K Baykov Aleksey E Sokolov Ludmila A Frank |
| Institution: | 1. Institute of Biophysics SB RAS, Federal Research Center KSC SB RAS, Krasnoyarsk, Russia;2. Siberian Federal University, Krasnoyarsk, Russia;3. Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk, Russia;4. Siberian Federal University, Krasnoyarsk, Russia
Kirensky Institute of Physics, Federal Research Center KSC SB RAS, Krasnoyarsk, Russia;5. Institute of Biophysics SB RAS, Federal Research Center KSC SB RAS, Krasnoyarsk, Russia Siberian Federal University, Krasnoyarsk, Russia |
| Abstract: | Bioluminescent solid-phase analysis was proposed to monitor the selection process and to determine binding characteristics of the aptamer–target complexes during design and development of the specific aptamers. The assay involves Ca2+-regulated photoprotein obelin as a simple, sensitive and fast reporter. Applicability and the prospects of the approach were exemplified by identification of DNA aptamers to cardiac troponin I, a highly specific early biomarker for acute myocardial infarction. Two structurally different aptamers specific to various epitopes of troponin I were obtained and then tested in a model bioluminescent assay. |
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