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An In Vitro Selection Platform to Identify Multiple Aptamers against Multiple Cell-Surface Markers Using
Nicole B Williams1, Sana Batool2, Hasan E Zumrut3
1Ph.D. Program in Molecular, Cellular and Developmental Biology, CUNY Graduate Center, The City University of New York, 365 Fifth Avenue, New York, New York 10016, United States.
Biochemistry
|July 13, 2022
Summary
This study expands ligand-guided selection (LIGS) to discover aptamers targeting multiple cell-surface receptors simultaneously. This multiplexed approach enables the development of novel molecular tools for complex biological targets.
Area of Science:
- Biotechnology
- Molecular Biology
- Biochemistry
Background:
- Aptamer ligand discovery is crucial for developing molecular tools, but cell-surface receptors present challenges.
- Systematic evolution of ligands by exponential enrichment (SELEX) variants aim to improve aptamer screening.
- Previous ligand-guided selection (LIGS) focused on single dominant cell-surface receptors.
Purpose of the Study:
- To expand LIGS for identifying aptamers against multiple cell-surface receptors on the same cell.
- To establish a multiplexed ligand discovery platform using SELEX-based methods.
- To generate specific aptamers against CD19 and CD20 expressed on Toledo cells.
Main Methods:
- Developed and applied a multi-target LIGS strategy.
- Evolved aptamer families against Toledo cells expressing CD19 and CD20.
- Utilized monoclonal antibodies for selective partitioning of aptamers against specific targets.
- Performed biochemical characterization of evolved aptamers.
Main Results:
- Successfully identified multiple aptamer families against Toledo cells.
- Isolated two specific aptamers against CD19 and two against CD20 with high affinity.
- Demonstrated the feasibility of targeting multiple receptors on a single cell surface.
Conclusions:
- Multi-target LIGS is a viable combinatorial approach for nucleic acid library screening.
- This method enables the generation of multiple aptamers against multiple membrane receptors.
- The developed platform facilitates the creation of advanced molecular tools for complex cellular targets.
