Abstract
The ability of aptamers to recognize a variety of different molecules has fueled their emergence as recognition agents to probe complex media and cells. Many detection strategies require aptamer binding to its target to result in a dramatic change in structure, typically from an unfolded to a folded state. Here, we report a strategy based on forced intercalation (FIT) that increases the scope of aptamer recognition by transducing subtle changes in aptamer structures into fluorescent readouts. By screening a library of green-fluorescent FIT-aptamers whose design is guided by computational modeling, we could identify hits that sense steroids like dehydroepiandrosterone sulfate (DHEAS) down to 1.3 μM with no loss in binding affinity compared to the unmodified aptamer. This enabled us to study DHEAS in clinical serum samples with several advantages over gold standard methods, including rapid readout (<30 min), simple instrumentation (plate-reader), and low sample volumes (10 μL).
| Original language | English (US) |
|---|---|
| Pages (from-to) | 15260-15265 |
| Number of pages | 6 |
| Journal | Angewandte Chemie - International Edition |
| Volume | 60 |
| Issue number | 28 |
| DOIs | |
| State | Published - Jul 5 2021 |
Funding
This material is based on research sponsored by Air Force Research Laboratory under agreement number FA8650-15-2-5518. The U.S. Government is authorized to reproduce and distribute reprints for Governmental purposes notwithstanding any copyright notation thereon. The views and conclusions contained herein are those of the authors and should not be interpreted as necessarily representing the official policies or endorsements, either expressed or implied, of Air Force Research Laboratory or the U.S. Government. This material is also based upon work supported by the Air Force Office of Scientific Research awards FA9550-16-1-0150 and FA9550-17-1-0348, the National Science Foundation grant CHE-1709888, and the Sherman Fairchild Foundation, Inc. This work made use of the IMSERC MS facility at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-2025633), the State of Illinois, and the International Institute for Nanotechnology (IIN). This work made use of the Keck Biophysics Facility supported by the NCI CCSG P30 CA060553 grant awarded to the Robert H Lurie Comprehensive Cancer. S.B.E. was supported in part by the Chicago Cancer Baseball Charities and the H Foundation at the Lurie Cancer Center of Northwestern University. This material is based on research sponsored by Air Force Research Laboratory under agreement number FA8650‐15‐2‐5518. The U.S. Government is authorized to reproduce and distribute reprints for Governmental purposes notwithstanding any copyright notation thereon. The views and conclusions contained herein are those of the authors and should not be interpreted as necessarily representing the official policies or endorsements, either expressed or implied, of Air Force Research Laboratory or the U.S. Government. This material is also based upon work supported by the Air Force Office of Scientific Research awards FA9550‐16‐1‐0150 and FA9550‐17‐1‐0348, the National Science Foundation grant CHE‐1709888, and the Sherman Fairchild Foundation, Inc. This work made use of the IMSERC MS facility at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS‐2025633), the State of Illinois, and the International Institute for Nanotechnology (IIN). This work made use of the Keck Biophysics Facility supported by the NCI CCSG P30 CA060553 grant awarded to the Robert H Lurie Comprehensive Cancer. S.B.E. was supported in part by the Chicago Cancer Baseball Charities and the H Foundation at the Lurie Cancer Center of Northwestern University.
Keywords
- DNA
- aptamers
- detection
- forced intercalation
- steroids
ASJC Scopus subject areas
- Catalysis
- General Chemistry