A general double library SELEX strategy for aptamer selection using unmodified nonimmobilized targets
- 735 Downloads
Aptamer discovery for unmodified nonimmobilized targets has been constantly presenting itself as a significant challenge to the research community. We demonstrate here a novel double library (DL) SELEX strategy and its usefulness and generality toward discovering both ssDNA- and RNA-based aptamers with nanomolar binding affinities toward unmodified targets of both small (e.g., doxycycline) and large (e.g., VEGF165) sizes. The same selection strategy further allows for concurrent selection of an aptamer pair, recognizing discrete epitopes on the same protein, from the same selection cycles for the sandwich aptamer pair-based biosensor development (e.g., one aptamer for the recognition and the other for the signal transduction). These results establish the DL-SELEX method developed here as a valuable and highly accessible selection strategy for aptamer discovery, especially when chemical modifications of target molecules are not preferred or simply impossible.
KeywordsAptamer SELEX Nucleic acids Nonimmobilization Label-free
This work was supported by the Institute of Bioengineering and Nanotechnology (Biomedical Research Council, Agency for Science, Technology and Research, Singapore).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 36.Xiao SJ, Hu PP, Wu XD, Zou YL, Chen LQ, Peng L, et al. Sensitive discrimination and detection of prion disease-associated isoform with a dual-aptamer strategy by developing a sandwich structure of magnetic microparticles and quantum dots. Anal Chem. 2010;82(23):9736–42. doi: 10.1021/ac101865s.CrossRefGoogle Scholar