Protein-environment-sensitive computational epitope accessibility analysis from antibody dose-response data
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arXiv
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| Main Authors: | , , , , |
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| Format: | Preprint |
| Published: |
2024
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| _version_ | 1866910517956706304 |
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| author | Tschimmel, Dominik Saeed, Momina Milani, Maria Waldherr, Steffen Hucho, Tim |
| author_facet | Tschimmel, Dominik Saeed, Momina Milani, Maria Waldherr, Steffen Hucho, Tim |
| contents | Antibodies are widely used in life-sciences and medical therapy. Yet, broadly applicable methods are missing to determine, in the biological system of choice, antibody specificity and its quantitative contribution to e.g. immunofluorescence stainings. Thereby, antibody-based data often needs to be seen with caution. Here, we present a simple-to-use approach to characterize and quantify antibody binding properties directly in the system of choice. We determine an epitope accessibility distribution in the system of interest based on a computational analysis of antibody-dilution immunofluorescence stainings. This allows the selection of specific antibodies, the choice of a dilution to maximize signal-specificity, and an improvement of signal quantification. It further expands the scope of antibody-based imaging to detect changes of the subcellular nano-environment and allows for antibody multiplexing. |
| format | Preprint |
| id |
arxiv_https___arxiv_org_abs_2407_06052 |
| institution | arXiv |
| publishDate | 2024 |
| record_format | arxiv |
| spellingShingle | Protein-environment-sensitive computational epitope accessibility analysis from antibody dose-response data Tschimmel, Dominik Saeed, Momina Milani, Maria Waldherr, Steffen Hucho, Tim Quantitative Methods Antibodies are widely used in life-sciences and medical therapy. Yet, broadly applicable methods are missing to determine, in the biological system of choice, antibody specificity and its quantitative contribution to e.g. immunofluorescence stainings. Thereby, antibody-based data often needs to be seen with caution. Here, we present a simple-to-use approach to characterize and quantify antibody binding properties directly in the system of choice. We determine an epitope accessibility distribution in the system of interest based on a computational analysis of antibody-dilution immunofluorescence stainings. This allows the selection of specific antibodies, the choice of a dilution to maximize signal-specificity, and an improvement of signal quantification. It further expands the scope of antibody-based imaging to detect changes of the subcellular nano-environment and allows for antibody multiplexing. |
| title | Protein-environment-sensitive computational epitope accessibility analysis from antibody dose-response data |
| topic | Quantitative Methods |
| url | https://arxiv.org/abs/2407.06052 |