Journal
ANALYTICAL BIOCHEMISTRY
Volume 658, Issue -, Pages -Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2022.114849
Keywords
Bispecific aptamer; Capture-SELEX; Malachite green; Leucomalachite green
Funding
- National Natural Science Foundation of China [31602126]
- Foundation of Educational Department of Jilin Province [JJKH20180901KJ]
- Youth Science Foundation [602017020]
- Application-oriented Foundation of Yanbian University [602021007]
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The bispecific aptamers against MG and LMG were isolated using Capture-SELEX, and their specificity and affinity were confirmed. This study provides novel probes for the development of multi-residues rapid detection and opportunities for multiple target aptamer discovery.
In order to develop multi-residues rapid detection, the bispecific aptamers against malachite green (MG) and leucomalachite green (LMG) were isolated by the capture systematic evolution of ligands by exponential enrichment (Capture-SELEX). After thirteen rounds of selection, the enriched ssDNA pools were sent for high -throughput sequencing. Nine aptamer candidates (A1-A9) were picked out to test their specificity by gold nanoparticles (AuNPs) colorimetric assay. Three aptamers (A2, A3, A5) with good selectivity were truncated to verify their affinity by fluorescence assay. Finally, three truncated aptamers (A2-a, A3-a, A5-a) with bispecificity and high affinity were identified. For LMG, the dissociation constant (Kd) of them were 8.4 +/- 0.8 nM, 8.2 +/- 1.2 nM, and 13.7 +/- 1.4 nM, respectively. For MG, Kd of them were 3.4 +/- 0.3 mu M, 2.3 +/- 0.2 mu M, 3.0 +/- 0.2 mu M. Among them, A3-a is the best. Our work will provide novel probes for the development of multi-residues rapid detection as well as opportunities for multiple target aptamer discovery.
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