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MALDI-IHC-Guided In-Depth Spatial Proteomics: Targeted and Untargeted MSI Combined
[Image: see text] Recently, a novel technology was published, utilizing the strengths of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) and immunohistochemistry (IHC), achieving highly multiplexed, targeted imaging of biomolecules in tissue. This new technique, c...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9893213/ https://www.ncbi.nlm.nih.gov/pubmed/36638208 http://dx.doi.org/10.1021/acs.analchem.2c04220 |
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author | Claes, Britt S. R. Krestensen, Kasper K. Yagnik, Gargey Grgic, Andrej Kuik, Christel Lim, Mark J. Rothschild, Kenneth J. Vandenbosch, Michiel Heeren, Ron M. A. |
author_facet | Claes, Britt S. R. Krestensen, Kasper K. Yagnik, Gargey Grgic, Andrej Kuik, Christel Lim, Mark J. Rothschild, Kenneth J. Vandenbosch, Michiel Heeren, Ron M. A. |
author_sort | Claes, Britt S. R. |
collection | PubMed |
description | [Image: see text] Recently, a novel technology was published, utilizing the strengths of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) and immunohistochemistry (IHC), achieving highly multiplexed, targeted imaging of biomolecules in tissue. This new technique, called MALDI-IHC, opened up workflows to target molecules of interest using MALDI-MSI that are usually targeted by standard IHC. In this paper, the utility of targeted MALDI-IHC and its complementarity with untargeted on-tissue bottom-up spatial proteomics is explored using breast cancer tissue. Furthermore, the MALDI-2 effect was investigated and demonstrated to improve MALDI-IHC. Formalin-fixed paraffin-embedded (FFPE) human breast cancer tissue sections were stained for multiplex MALDI-IHC with six photocleavable mass-tagged (PC-MT) antibodies constituting a breast cancer antibody panel (CD20, actin-αSM, HER2, CD68, vimentin, and panCK). K-means spatial clusters were created based on the MALDI-IHC images and cut out using laser-capture microdissection (LMD) for further untargeted LC-MS-based bottom-up proteomics analyses. Numerous peptides could be tentatively assigned to multiple proteins, of which three proteins were also part of the antibody panel (vimentin, keratins, and actin). Post-ionization with MALDI-2 showed an increased intensity of the PC-MTs and suggests options for the development of new mass-tags. Although the on-tissue digestion covered a wider range of proteins, the MALDI-IHC allowed for easy and straightforward identification of proteins that were not detected in untargeted approaches. The combination of the multiplexed MALDI-IHC with image-guided proteomics showed great potential to further investigate diseases by providing complementary information from the same tissue section and without the need for customized instrumentation. |
format | Online Article Text |
id | pubmed-9893213 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-98932132023-02-03 MALDI-IHC-Guided In-Depth Spatial Proteomics: Targeted and Untargeted MSI Combined Claes, Britt S. R. Krestensen, Kasper K. Yagnik, Gargey Grgic, Andrej Kuik, Christel Lim, Mark J. Rothschild, Kenneth J. Vandenbosch, Michiel Heeren, Ron M. A. Anal Chem [Image: see text] Recently, a novel technology was published, utilizing the strengths of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) and immunohistochemistry (IHC), achieving highly multiplexed, targeted imaging of biomolecules in tissue. This new technique, called MALDI-IHC, opened up workflows to target molecules of interest using MALDI-MSI that are usually targeted by standard IHC. In this paper, the utility of targeted MALDI-IHC and its complementarity with untargeted on-tissue bottom-up spatial proteomics is explored using breast cancer tissue. Furthermore, the MALDI-2 effect was investigated and demonstrated to improve MALDI-IHC. Formalin-fixed paraffin-embedded (FFPE) human breast cancer tissue sections were stained for multiplex MALDI-IHC with six photocleavable mass-tagged (PC-MT) antibodies constituting a breast cancer antibody panel (CD20, actin-αSM, HER2, CD68, vimentin, and panCK). K-means spatial clusters were created based on the MALDI-IHC images and cut out using laser-capture microdissection (LMD) for further untargeted LC-MS-based bottom-up proteomics analyses. Numerous peptides could be tentatively assigned to multiple proteins, of which three proteins were also part of the antibody panel (vimentin, keratins, and actin). Post-ionization with MALDI-2 showed an increased intensity of the PC-MTs and suggests options for the development of new mass-tags. Although the on-tissue digestion covered a wider range of proteins, the MALDI-IHC allowed for easy and straightforward identification of proteins that were not detected in untargeted approaches. The combination of the multiplexed MALDI-IHC with image-guided proteomics showed great potential to further investigate diseases by providing complementary information from the same tissue section and without the need for customized instrumentation. American Chemical Society 2023-01-13 /pmc/articles/PMC9893213/ /pubmed/36638208 http://dx.doi.org/10.1021/acs.analchem.2c04220 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Claes, Britt S. R. Krestensen, Kasper K. Yagnik, Gargey Grgic, Andrej Kuik, Christel Lim, Mark J. Rothschild, Kenneth J. Vandenbosch, Michiel Heeren, Ron M. A. MALDI-IHC-Guided In-Depth Spatial Proteomics: Targeted and Untargeted MSI Combined |
title | MALDI-IHC-Guided
In-Depth Spatial Proteomics: Targeted
and Untargeted MSI Combined |
title_full | MALDI-IHC-Guided
In-Depth Spatial Proteomics: Targeted
and Untargeted MSI Combined |
title_fullStr | MALDI-IHC-Guided
In-Depth Spatial Proteomics: Targeted
and Untargeted MSI Combined |
title_full_unstemmed | MALDI-IHC-Guided
In-Depth Spatial Proteomics: Targeted
and Untargeted MSI Combined |
title_short | MALDI-IHC-Guided
In-Depth Spatial Proteomics: Targeted
and Untargeted MSI Combined |
title_sort | maldi-ihc-guided
in-depth spatial proteomics: targeted
and untargeted msi combined |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9893213/ https://www.ncbi.nlm.nih.gov/pubmed/36638208 http://dx.doi.org/10.1021/acs.analchem.2c04220 |
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