Institute of Biophysics, Russian Academy of Sciences SB, Krasnoyarsk 660036, Russia.
Anal Bioanal Chem. 2011 Nov;401(8):2573-9. doi: 10.1007/s00216-011-5343-2. Epub 2011 Sep 4.
The recombinant Ca(2+)-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.
来自海肾的重组 Ca(2+)-触发腔肠素结合蛋白 (CBP) 被研究为用于体外分析应用的生物特异性标记分子。该蛋白在冷冻状态的溶液中表现稳定，在加热和化学修饰下也表现稳定。获得了与生物素、寡核苷酸和蛋白质的缀合物，并将其作为生物特异性分子应用于固相微分析中。在存在钙的情况下，使用完整的（未进行任何修饰）海肾荧光素酶进行 CBP 检测，发现检测限为 75 飞摩尔。模型实验表明，这种方法具有很大的前景，特别是在多分析系统的开发方面。
