Synthesis, Characterization, and Applications of a Superior Dendrimer-Based Polymer for Multiplexed Ion Beam Imaging Time-of-Flight Analysis
- 23 June 2023
- journal article
- research article
- Published by American Chemical Society (ACS) in Biomacromolecules
- Vol. 24 (7) , 3105-3114
Abstract
High-dimensional single-cell mass spectrometric imaging techniques such as multiplexed ion beam imaging by time-of-flight mass spectrometry (MIBI-TOF), imaging mass cytometry (IMC), and flow cytometry-based CyTOF utilize antibodies conjugated to linear metal-chelating polymers. Here, we report on the synthesis and characterization of a dendrimer-based polymer and its utilization in tissue imaging using MIBI-TOF. We compared the staining performance in FFPE tissue of antibodies for lineage-specific immune proteins (CD20, CD3, CD45, FoxP3) that were conjugated with dendrimer or linear polymer. Staining of serial tissue sections with dendron-conjugated and linear-polymer-conjugated antibodies revealed comparable avidities of dendrons and linear polymers with log2 (ratio of mean positive pixel intensity of staining for linear polymers to dendrons) within the range ±0.25. Interestingly, dendron-conjugated antibodies were observed to have some advantages over linear polymer-conjugated antibodies. For example, tissue staining of a nuclear protein, FoxP3 with dendron-conjugated antibodies showed notably less background staining than that of linear-polymer-conjugated antibodies. Additionally, dendron-conjugated antibodies did not exhibit off-target cytosolic binding in neural tissue typically observed when using linear polymer conjugates. Taken together, this work provides a versatile framework for using third-generation dendron-conjugated antibodies with improved staining over conventional linear polymers.Keywords
Funding Information
- U.S. Department of Health and Human Services (1R01CA24063801A1, 1U24CA22430901, 5R01CA22952904, 5U54CA20997105, 5UH3CA24663303)
- U.S. Department of Health and Human Services (5DP5OD01982205)
- Wellcome Trust
- Bill and Melinda Gates Foundation
- U.S. Department of War (W81XWH2110143)
- U.S. Department of Health and Human Services (5R01AG05628705, 5R01AG05791504, 5R01AG06827902)
- Breast Cancer Research Foundation
- Parker Institute for Cancer Immunotherapy
- Cancer Research Institute
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