ArticleCell reports methods2022
Engineered fast-dissociating antibody fragments for multiplexed super-resolution microscopy.
Article in Cell reports methods, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 13 citations in OpenAlex.
- Dynamic Binder Exchange Improves Protein Labeling Efficiency in DNA-PAINT up to 15-Fold.Angewandte Chemie (International ed. in English) · 2026Article
- Color mapping of multitarget protein clusters.Structure (London, England : 1993) · 2025Article
- Laminar organization of molecular complexes in a clathrin coat revealed by nanoscale protein colocalization.Structure (London, England : 1993) · 2025Article
- IntAct: A nondisruptive internal tagging strategy to study the organization and function of actin isoforms.PLoS biology · 2024Article
- Single-Molecule Localization Microscopy Using Time-Lapse Imaging of Single-Antibody Labeling.Current protocols · 2023Article
- Beyond DNA: new probes for PAINT super-resolution microscopy.Chemical communications (Cambridge, England) · 2023Review
- Superresolution Imaging with Single-Antibody Labeling.Bioconjugate chemistry · 2023Article
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Authors and funding
9 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Image reconstruction by integrating exchangeable single-molecule localization (IRIS) achieves multiplexed super-resolution imaging by high-density labeling with fast exchangeable fluorescent probes. However, previous methods to develop probes for individual targets required a great amount of time and effort. Here, we introduce a method for generating recombinant IRIS probes with a new mutagenesis strategy that can be widely applied to existing antibody sequences. Several conserved tyrosine residues at the base of complementarity-determining regions were identified as candidate sites for site-directed mutagenesis. With a high probability, mutations at candidate sites accelerated the off rate of recombinant antibody-based probes without compromising specific binding. We were able to develop IRIS probes from five monoclonal antibodies and three single-domain antibodies. We demonstrate multiplexed localization of endogenous proteins in primary neurons that visualizes small synaptic connections with high binding density. It is now practically feasible to generate fast-dissociating fluorescent probes for multitarget super-resolution imaging.
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