Computational epitope heterogeneity analysis in immunostainings from antibody-dilution series
- Author(s)
- Dominik Tschimmel, Momina Saeed, Maria Milani, Steffen Waldherr, Tim Hucho
- Abstract
Antibodies are widely used in life sciences and medical therapy. Broadly applicable methods to determine epitope heterogeneity in immunostaining systems are missing. Here, we present a simple-to-use approach to characterize and quantify antibody binding properties that constitute the staining directly in the system of choice. We determine an epitope heterogeneity on the basis of a computational analysis of antibody-dilution immunofluorescence stainings. This allows us to choose signal-specificity maximizing dilutions and to improve signal quantification. Furthermore, the computational analysis provides approaches to obtain a single-channel antibody multiplexing. Our approach could help improving immunostainings in many laboratories by guiding the choice of antibody dilution, by increasing the possibility of antibody-multiplexing in the same color-channel and by allowing for the analysis of binding targets of multi-specific antibodies.
- Organisation(s)
- Functional and Evolutionary Ecology
- External organisation(s)
- University Hospital Cologne
- Journal
- Communications Biology
- Volume
- 9
- ISSN
- 2399-3642
- DOI
- https://doi.org/10.1038/s42003-026-09517-x
- Publication date
- 2026
- Peer reviewed
- Yes
- Austrian Fields of Science 2012
- 106023 Molecular biology, 106045 Theoretical biology
- ASJC Scopus subject areas
- Medicine (miscellaneous), General Biochemistry,Genetics and Molecular Biology, General Agricultural and Biological Sciences
- Portal url
- https://ucrisportal.univie.ac.at/en/publications/ae1869aa-f50b-4db6-9eed-646dd46bd5a2