Rapid and accurate quantification of biomolecules in complex biological samples is essential in academic and practical applications. While traditional analytical and diagnostic technologies have exploited enzymes and antibodies, short synthetic nucleic acid sequences known as aptamers have recently emerged as an attractive alternative [1–3]. Their small size, synthetic nature, and remarkable thermal and structural stability make them promising candidates for developing portable real-time devices and biosensors [4].
In our laboratory, we have established the Aptamer Discovery and Application Platform (ADAP) that enables simultaneous selection of DNA aptamers against diverse ligands, such as proteins, small molecules and protein:small molecule complexes. The developed aptamers are then used for the construction of biosensing and analytical systems.
Figure 1. Overview of ADAP technology for multiplexed aptamer selection. The aptamer selection process involves the Capture-SELEX method (A), utilising a native target and immobilised DNA library. The multiplexed semi-automated SELEX is facilitated by a programmable magnetic particle processing platform (B). Following aptamer enrichment, the library undergoes NGS sequencing (C), and subsequent analysis and characterisation are performed using various methods (D). Post-SELEX optimisation of aptamers (E) is carried out to fine-tune them for sensing applications, e.g., fluorescent assays (F).
1. Yang,K., Mitchell,N.M., Banerjee,S., Cheng,Z., Taylor,S., Kostic,A.M., Wong,I., Sajjath,S., Zhang,Y., Stevens,J., et al. (2023) A functional group-guided approach to aptamers for small molecules. Science, 380, 942–948.
2. Yu,H., Alkhamis,O., Canoura,J., Liu,Y. and Xiao,Y. (2021) Advances and Challenges in Small-Molecule DNA Aptamer Isolation, Characterization, and Sensor Development. Angew. Chemie – Int. Ed., 60, 16800–16823.
3. Yang,L.F., Ling,M., Kacherovsky,N. and Pun,S.H. (2023) Aptamers 101: aptamer discovery and in vitro applications in biosensors and separations. Chem. Sci., 14, 4961–4978.
4. Downs,A.M., Gerson,J., Leung,K.K., Honeywell,K.M., Kippin,T. and Plaxco,K.W. (2022) Improved calibration of electrochemical aptamer-based sensors. Sci. Rep., 12, 1–10.