ArticleProceedings of the National Academy of Sciences of the United States of America2018
Streamlined circular proximity ligation assay provides high stringency and compatibility with low-affinity antibodies.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 31 citations in OpenAlex.
- In situ tracking of glycoRNAs on single-cell surface to reveal RNA heterogeneity and transport mechanism.Nucleic acids research · 2026Article
- Aptamer-based approaches for sensitive detection and epitope mapping of SARS-CoV-2 spike protein.Molecular therapy. Nucleic acids · 2026Article
- Mapping GlycoRNAs on an Exosomal Surface.Journal of the American Chemical Society · 2026Article
- Proximity Ligation Assay: From a Foundational Principle to a Versatile Platform for Molecular and Translational Research.Biomolecules · 2025Review
- Spatially resolved subcellular protein-protein interactomics in drug-perturbed lung-cancer cultures and tissues.Nature biomedical engineering · 2025Article
- Single-cell imaging of N4-acetylcytidine-modified RNA using fluorine metabolic labeling mediated proximity ligation assay.Nucleic acids research · 2025Article
- Proximity-activated DNA scanning encoded sequencing for massive access to membrane proteins nanoscale organization.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- CHD4 and SMYD1 repress common transcriptional programs in the developing heart.Development (Cambridge, England) · 2024Article
- Spatial imaging of glycoRNA in single cells with ARPLA.Nature biotechnology · 2024Article
- Visualization of Individual RNA Molecules by Proximity Ligation-Based Chromogenic In Situ Hybridization Assay.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Extending the dynamic range of biomarker quantification through molecular equalization.Nature communications · 2023Article
- Assay methods based on proximity-enhanced reactions for detecting non-nucleic acid molecules.Frontiers in bioengineering and biotechnology · 2023Review
- CHD4 is recruited by GATA4 and NKX2-5 to repress noncardiac gene programs in the developing heart.Genes & development · 2022Article
- ER-mitochondria contact sites in neurodegeneration: genetic screening approaches to investigate novel disease mechanisms.Cell death and differentiation · 2021Review
- Nucleic-Acid Driven Cooperative Bioassays Using Probe Proximity or Split-Probe Techniques.Analytical chemistry · 2021Review
- Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction.Frontiers in cell and developmental biology · 2021Review
- Molecular and Biochemical Techniques for Deciphering p53-MDM2 Regulatory Mechanisms.Biomolecules · 2020Review
- High-Fidelity Nanopore Sequencing of Ultra-Short DNA Targets.Analytical chemistry · 2019Article
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Authors and funding
5 authors at 1 institution in 1 country.
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Abstract
Proximity ligation assay (PLA) is a powerful tool for quantitative detection of protein biomarkers in biological fluids and tissues. Here, we present the circular proximity ligation assay (c-PLA), a highly specific protein detection method that outperforms traditional PLA in stringency, ease of use, and compatibility with low-affinity reagents. In c-PLA, two proximity probes bind to an analyte, providing a scaffolding that positions two free oligonucleotides such that they can be ligated into a circular DNA molecule. This assay format stabilizes antigen proximity probe complexes and enhances stringency by reducing the probability of random background ligation events. Circle formation also increases selectivity, since the uncircularized DNA can be removed enzymatically. We compare this method with traditional PLA on several biomarkers and show that the higher stringency for c-PLA improves reproducibility and enhances sensitivity in both buffer and human plasma. The limit of detection ranges from femtomolar to nanomolar concentrations for both methods. Kinetic analyses using surface plasmon resonance (SPR) and biolayer interferometry (BLI) reveal that the variation in limit of detection is due to the variation in antibody affinity and that c-PLA outperforms traditional PLA for low-affinity antibodies. The lower background signal can be used to increase proximity probe concentration while maintaining a high signal-to-noise ratio, thereby enabling the use of low-affinity reagents in a homogeneous assay format. We anticipate that the advantages of c-PLA will be useful in a variety of clinical protein detection applications where high-affinity reagents are lacking.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.