Journal of Siberian Federal University. Biology / Bioluminescent Binding Microassay Using Aptamers as Biospecific Elements

Full text (.pdf)
Issue
Journal of Siberian Federal University. Biology. 2019 12 (3)
Authors
Frank, Ludmila A.; Bashmakova, Eugenia E.; Goncharova, Natalia S.; Krasitskaya, Vasilisa V.
Contact information
Frank, Ludmila A.: Institute of Biophysics SB RAS Federal Research Center “Krasnoyarsk Science Center SB RAS” 50/50 Akademgorodok, Krasnoyarsk, 660036, Russia; Siberian Federal University 79 Svobodny, Krasnoyarsk, 660041, Russia; ; ORCID: 0000-0003-4462-1944; Bashmakova, Eugenia E.: Institute of Biophysics SB RAS Federal Research Center “Krasnoyarsk Science Center SB RAS” 50/50 Akademgorodok, Krasnoyarsk, 660036, Russia; ruORCID: 0000-0002-8951-8599; Goncharova, Natalia S.: Institute of Biophysics SB RAS Federal Research Center “Krasnoyarsk Science Center SB RAS” 50/50 Akademgorodok, Krasnoyarsk, 660036, Russia; Siberian Federal University 79 Svobodny, Krasnoyarsk, 660041, Russia; Krasitskaya, Vasilisa V.: Institute of Biophysics SB RAS Federal Research Center “Krasnoyarsk Science Center SB RAS” 50/50 Akademgorodok, Krasnoyarsk, 660036, Russia; ORCID: 0000-0001-7018-2581
Keywords
aptamers; Ca2+-regulated photoprotein obelin; bioluminescent microassay
Abstract

High specificity is an important requirement for an analytical system aimed at identifying a specific molecular target. Traditionally, antibodies, haptens and some other molecules are used for this purpose. Recently, aptamers were proposed as biospecific elements. Aptamers are short single-stranded oligonucleotides with a unique spatial structure that enables them to recognize target molecules and bind to them. Aptamers are obtained from synthetic random DNA(RNA)-libraries, a pool of oligonucleotides of the same length with different base sequences (1014-1015 variants), by selecting the oligonucleotides that are capable of specific binding to a given target. Aptamers are stable molecules with high affinity and specificity; they can be developed for any target, including toxic and nonimmunogenic ones; and they can be easily synthesized chemically. Due to these useful qualities, aptamers are often considered to be an alternative to antibodies. This paper describes the use of aptamer sensors and a highly sensitive bioluminescent reporter, the Ca2+-regulated photoprotein obelin, for the detection of diagnostically important targets in the blood of patients. Additionally, obelin was successfully applied as a reporter in the process of obtaining aptamers. A proposed bioluminescent solid-phase assay enables the enrichment of the oligonucleotide library with target-specific oligonucleotides to be monitored rapidly, the affinity of individual aptamers and their shortened variants to be evaluated and the relative position of the aptamers on the target molecule to be determined. The results of the studies reviewed in this paper open promising avenues for developing analytical systems that include highly specific aptamer sensing, as well as highly sensitive detection based on bioluminescent reporter proteins

Pages
244-252
DOI
10.17516/1997-1389-0298
Paper at repository of SibFU
https://elib.sfu-kras.ru/handle/2311/125595

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