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The Power of Old Hats: Rediscovering Inosine-EpPCR to Create Starting Libraries for Whole-Cell-SELEX
Grigory Bolotnikov1, Ann-Kathrin Kissmann1, Daniel Gruber1
1Institute of Pharmaceutical Biotechnology, Ulm University, 89081 Ulm, Germany.
Biosensors
|July 25, 2025
Summary
Inosine-mediated error-prone PCR (epPCR) efficiently generates aptamer libraries. This cost-effective method simplifies aptamer development and SELEX campaigns, enabling functional selections against diverse targets.
Area of Science:
- Biotechnology
- Molecular Biology
- Genomics
Background:
- Aptamer library generation is crucial for SELEX.
- Traditional methods can be costly and complex.
- Inosine-mediated error-prone PCR (epPCR) is an underutilized technique.
Purpose of the Study:
- To demonstrate the efficacy of epPCR for aptamer library creation.
- To show epPCR suitability for initiating whole-cell SELEX.
- To establish epPCR as a robust and cost-effective method.
Main Methods:
- Utilized inosine to introduce targeted mutations via epPCR.
- Generated aptamer libraries from a single oligonucleotide.
- Applied epPCR-derived libraries to whole-cell SELEX against ten diverse targets.
Main Results:
- epPCR successfully generated diverse aptamer libraries.
- Enriched aptamers displayed novel binding specificities.
- The method proved effective for functional aptamer selection.
Conclusions:
- epPCR is a powerful, cost-effective strategy for aptamer library generation.
- This approach lowers the barrier for initiating SELEX campaigns.
- Revisiting established techniques like epPCR addresses modern aptamer development challenges.
