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Developing a Toolbox of Antibodies Validated for Array Tomography-Based Imaging of Brain Synapses
Kristina D Micheva1, Belvin Gong2, Forrest Collman3
1Department of Cellular and Molecular Physiology, Stanford School of Medicine, Stanford, CA.
Biorxiv : the Preprint Server for Biology
|July 10, 2023
Summary
Researchers developed a new screening method to find antibodies for array tomography (AT), a microscopy technique. This approach efficiently identifies suitable antibodies, improving proteomic analysis in neuroscience research.
Area of Science:
- Neuroscience
- Cell Biology
- Microscopy
Background:
- Antibody performance is application-specific, requiring extensive validation for new uses.
- Array tomography (AT) enables high-dimensional proteomic analysis but needs validated antibodies.
Approach:
- Developed a novel, application-specific proxy screening step for identifying AT-compatible antibodies.
- Created a heterologous cell-based assay mimicking AT conditions (fixation, embedding) to predict antibody binding.
- Integrated this assay into monoclonal antibody generation for AT applications.
Key Points:
- The proxy screening simplifies antibody selection and has high predictive value for AT suitability.
- A database of AT-validated antibodies for neuroscience was established.
- Validated antibodies show high success rates for post-embedding applications like immunogold electron microscopy.
Conclusions:
- This screening strategy efficiently identifies candidate antibodies for array tomography.
- A growing toolbox of AT-compatible antibodies will enhance the utility of this imaging technique.
- The approach facilitates proteomic analysis in neuroscience and beyond.
