Rational Truncation of Aptamer for Ultrasensitive Aptasensing of Chloramphenicol: Studies Using Bio-Layer Interferometry.

Richa, Sharma and Monali, Mukherjee and Praveena, Bhatt and Raghavarao, K. S. M. S. (2023) Rational Truncation of Aptamer for Ultrasensitive Aptasensing of Chloramphenicol: Studies Using Bio-Layer Interferometry. Biosensors, 13. p. 660.

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Abstract

Aptamers are an excellent choice for the selective detection of small molecules. However, the previously reported aptamer for chloramphenicol suffers from low affinity, probably as a result of steric hindrance due to its bulky nature (80 nucleotides) leading to lower sensitivity in analytical assays. The present work was aimed at improving this binding affinity by truncating the aptamer without compromising its stability and three-dimensional folding. Shorter aptamer sequences were designed by systematically removing bases from each or both ends of the original aptamer. Thermodynamic factors were evaluated computationally to provide insight into the stability and folding patterns of the modified aptamers. Binding affinities were evaluated using bio-layer interferometry. Among the eleven sequences generated, one aptamer was selected based on its low dissociation constant, length, and regression of model fitting with association and dissociation curves. The dissociation constant could be lowered by 86.93% by truncating 30 bases from the 30 end of the previously reported aptamer. The selected aptamer was used for the detection of chloramphenicol in honey samples, based on a visible color change upon the aggregation of gold nanospheres caused by aptamer desorption. The detection limit could be reduced 32.87 times (1.673 pg mL

Item Type:	Article
Uncontrolled Keywords:	antibiotics; affinity; biosensing; interferometry; nanoparticles
Subjects:	500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 11 Antibiotics Chemistry
Divisions:	Food Engineering Food Microbiology
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	07 Dec 2023 06:11
Last Modified:	07 Dec 2023 06:11
URI:	http://ir.cftri.res.in/id/eprint/16797

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