ArticleProceedings of the National Academy of Sciences of the United States of America2013
Quantitative selection and parallel characterization of aptamers.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers.
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Who cites it
50 citing papers in PubMed.
- Innovations in Aptamer Technology: SELEX To Intelligent Molecular Engineering and Clinical Translation.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2026Review
- High-affinity ssDNA aptamer and chemiluminescent aptasensor for TIMP-1 detection in human serum.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2025Article
- New Insights into Aptamers: An Alternative to Antibodies in the Detection of Molecular Biomarkers.International journal of molecular sciences · 2024Review
- Recent Aptamer-Based Biosensors for CdBiosensors · 2023Review
- High-throughput quantitative binding analysis of DNA aptamers using exonucleases.Nucleic acids research · 2023Article
- Polydopamine Nanoparticles Functionalized Electrochemical DNA Aptasensor for Serum Glycated Albumin Detection.International journal of molecular sciences · 2022Article
- De novo selected hACE2 mimics that integrate hotspot peptides with aptameric scaffolds for binding tolerance of SARS-CoV-2 variants.Science advances · 2022Article
- Efficient delivery of a DNA aptamer-based biosensor into plant cells for glucose sensing through thiol-mediated uptake.Science advances · 2022Article
- Characterizing Aptamers with Reconfigurable Chiral Plasmonic Assemblies.Langmuir : the ACS journal of surfaces and colloids · 2022Article
- ProtSeq: Toward high-throughput, single-molecule protein sequencing via amino acid conversion into DNA barcodes.iScience · 2022Article
- Molecular Recognition: Perspective and a New Approach.Sensors (Basel, Switzerland) · 2021Article
- Systematic Combination of Oligonucleotides and Synthetic Polymers for Advanced Therapeutic Applications.Macromolecular research · 2021Review
- Implementation of High-Throughput Sequencing (HTS) in Aptamer Selection Technology.International journal of molecular sciences · 2020Review
- Label-free profiling of DNA aptamer-small molecule binding using T5 exonuclease.Nucleic acids research · 2020Article
- High throughput sequencing of in vitro selections of mRNA-displayed peptides: data analysis and applications.Physical chemistry chemical physics : PCCP · 2020Review
- Rapid selection of aptamers based on protein microarray.RSC advances · 2019Article
- Poly-Target Selection Identifies Broad-Spectrum RNA Aptamers.Molecular therapy. Nucleic acids · 2018Article
- Molecular Application of Aptamers in the Diagnosis and Treatment of Cancer and Communicable Diseases.Pharmaceuticals (Basel, Switzerland) · 2018Review
- Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor.Journal of visualized experiments : JoVE · 2018Article
- Application of the Open qPCR Instrument for the in Vitro Selection of DNA Aptamers against Epidermal Growth Factor Receptor and Drosophila C Virus.ACS combinatorial science · 2018Article
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Authors and funding
10 authors.
Funding
Abstract
Aptamers are promising affinity reagents that are potentially well suited for high-throughput discovery, as they are chemically synthesized and discovered via completely in vitro selection processes. Recent advancements in selection, sequencing, and the use of modified bases have improved aptamer quality, but the overall process of aptamer generation remains laborious and low-throughput. This is because binding characterization remains a critical bottleneck, wherein the affinity and specificity of each candidate aptamer are measured individually in a serial manner. To accelerate aptamer discovery, we devised the Quantitative Parallel Aptamer Selection System (QPASS), which integrates microfluidic selection and next-generation sequencing with in situ-synthesized aptamer arrays, enabling simultaneous measurement of affinity and specificity for thousands of candidate aptamers in parallel. After using QPASS to select aptamers for the human cancer biomarker angiopoietin-2 (Ang2), we in situ synthesized arrays of the selected sequences and obtained equilibrium dissociation constants (Kd) for every aptamer in parallel. We thereby identified over a dozen high-affinity Ang2 aptamers, with Kd as low as 20.5 ± 7.3 nM. The same arrays enabled us to quantify binding specificity for these aptamers in parallel by comparing relative binding of differentially labeled target and nontarget proteins, and by measuring their binding affinity directly in complex samples such as undiluted serum. Finally, we show that QPASS offers a compelling avenue for exploring structure-function relationships for large numbers of aptamers in parallel by coupling array-based affinity measurements with next-generation sequencing data to identify nucleotides and motifs within the aptamer that critically affect Ang2 binding.
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