Selection and characterization of an ssDNA aptamer against thyroglobulin

Thyroglobulin (Tg) is a significant biomarker for the diagnose and postoperative monitoring of differentiated thyroid cancer, and its recognition is urgent due to the rising prevalence. In this study, an ssDNA aptamer against Tg was obtained by capillary electrophoresis-systematic evolution of ligands via exponential enrichment (CESELEX). Under the optimized conditions, the sub-library was enriched well through two selection rounds. After high-throughput sequencing, eight candidate sequences were picked out and their affinities towards Tg were observed not in accordance with the order of their frequencies, whereas sequence homology played a significant role in binding affinity. The high-affinity sequence Seq.T-2 with a dissociation constant (K-d) of 3.18 mu M was finally selected as the aptamer, and its affinity was confirmed qualitatively by gold nanoparticles colorimetric and quantitatively by thin film interferometry (K-d, 4.51 nM). Besides, molecular docking and dynamics simulation were performed for their binding sites prediction and affinity confirmation. Furthermore, the aptamer was applied for Tg detection, which delivered a detection limit of 5.0 nM as well as with good selectivity, and showed a good linear relationship within a wide range of 10 nM-6.4 mu M of Tg spiked into the serum matrix. This study first reported Tg's aptamer which also exhibited the potential in real applications.

Automated summary

In this study, a ssDNA aptamer against thyroglobulin (Tg) was successfully obtained using CESELEX, with sequence homology and high-throughput sequencing confirming the Kd values. The aptamer showed good performance in detecting Tg.