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Single-Molecule Mapping Landscape of Multivalent Antibody-DNA Framework Conjugates.
Qinglin Xia1,2, Mo Zhou1,2, Xia Liu3,4
1Division of Physical Biology, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China.
DNA nanostructures enable antibody complex assembly for cellular signaling and theranostics. This study maps antibody binding efficiencies on DNA origami, revealing how site distribution impacts conjugation, crucial for optimizing diagnostics and therapeutics.
Area of Science:
- Bioconjugation
- Nanotechnology
- Molecular Biology
Background:
- DNA nanostructures offer precise platforms for assembling multivalent antibody complexes.
- Evaluating protein conjugation efficiency on DNA templates is critical for applications in cellular signaling and theranostics.
- Heterogeneity in antibody attachment poses a challenge for predictable nanostructure function.
Purpose of the Study:
- To quantitatively assess antibody conjugation efficiency at different positions on DNA origami frameworks.
- To investigate the influence of docking site distribution on antibody binding patterns.
- To provide insights for optimizing DNA-based nanostructures for advanced applications.
Main Methods:
- Utilized atomic force microscopy (AFM) for single-molecule analysis of antibody coupling.
- Employed two-dimensional rectangular DNA origami frameworks with varying docking site distributions.
- Generated spatial maps of antibody binding efficiencies across DNA structures.
Main Results:
- Discrete docking sites (≥18 nm spacing) showed decreased antibody coupling efficiency from periphery to center.
- Continuous docking sites (∼10 nm spacing) exhibited higher central antibody coupling efficiency.
- Observed trends attributed to Coulombic repulsion, steric hindrance, and multivalent cooperative effects.
Conclusions:
- Developed a quantitative tool for evaluating protein-DNA framework conjugates.
- Demonstrated that docking site distribution significantly modulates antibody conjugation efficiency.
- Findings guide the optimization of DNA nanostructures for enhanced precision in diagnostics and therapeutics.
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