Journal
SENSORS AND ACTUATORS B-CHEMICAL
Volume 331, Issue -, Pages -Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2021.129442
Keywords
Aptamer truncation; Si nuclease digestion; PD-Ll; Signal dual amplification aptasensor
Funding
- National Key Research and Development Program of China [2018YFA0903000]
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Optimizing aptamer truncation can enhance specificity and robustness through rational design based on nuclease digestion, as demonstrated in the development of a highly sensitive fluorescence aptasensor for PD-L1 detection.
Truncation optimization of aptamers can improve both specificity and robustness. At present, aptamer truncation is mainly performed based on predictions from molecular docking simulations, but this usually requires tedious trial-and-error and may result in false predictions. Here, we propose a strategy based on digestion by S1 nuclease to rationally truncate the aptamer, using the PD-L1-binding aptamer Apt80 as an example. Due to steric hindrance, recognition and binding regions between Apt80 and PD-L1 are not digested by the enzyme. The truncated form, Apt38, shows higher affinity and larger conformational change after binding, when compared to Apt80. The truncated Apt38 was used as a platform for developing a signal dual amplification fluorescence aptasensor using targeted recycling assisted by exonuclease I and qRT-PCR analysis. This aptasensor exhibited a high sensitivity toward PD-L1 with a limit of detection of 0.076 ng/mL in buffer system and 0.3625 ng/mL in human serum. Owing to its high sensitivity, specificity, ease operation and low cost for detection of PD-L1, this aptasensor should be useful in assisting clinicians to evaluate the status of cancer patient and to decide whether inhibitor drugs are needed.
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