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Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies
Proteases are involved in the control of numerous physiological processes, and their dysregulation has been identified in a wide range of pathologies, including cancer. Protease activity is normally tightly regulated post-translationally and therefore cannot be accurately estimated based on mRNA or...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8471274/ https://www.ncbi.nlm.nih.gov/pubmed/34575469 http://dx.doi.org/10.3390/pharmaceutics13091390 |
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author | Howng, Bruce Winter, Michael B. LePage, Carol Popova, Irina Krimm, Michael Vasiljeva, Olga |
author_facet | Howng, Bruce Winter, Michael B. LePage, Carol Popova, Irina Krimm, Michael Vasiljeva, Olga |
author_sort | Howng, Bruce |
collection | PubMed |
description | Proteases are involved in the control of numerous physiological processes, and their dysregulation has been identified in a wide range of pathologies, including cancer. Protease activity is normally tightly regulated post-translationally and therefore cannot be accurately estimated based on mRNA or protein expression alone. While several types of zymography approaches to estimate protease activity exist, there remains a need for a robust and reliable technique to measure protease activity in biological tissues. We present a novel quantitative ex vivo zymography (QZ) technology based on Probody(®) therapeutics (Pb-Tx), a novel class of protease-activated cancer therapeutics that contain a substrate linker cleavable by tumor-associated proteases. This approach enables the measurement and comparison of protease activity in biological tissues via the detection of Pb-Tx activation. By exploiting substrate specificity and selectivity, cataloguing and differentiating protease activities is possible, with further refinement achieved using protease-specific inhibitors. Using the QZ assay and human tumor xenografts, patient tumor tissues, and patient plasma, we characterized protease activity in preclinical and clinical samples. The QZ assay offers the potential to increase our understanding of protease activity in tissues and inform diagnostic and therapeutic development for diseases, such as cancer, that are characterized by dysregulated proteolysis. |
format | Online Article Text |
id | pubmed-8471274 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-84712742021-09-27 Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies Howng, Bruce Winter, Michael B. LePage, Carol Popova, Irina Krimm, Michael Vasiljeva, Olga Pharmaceutics Article Proteases are involved in the control of numerous physiological processes, and their dysregulation has been identified in a wide range of pathologies, including cancer. Protease activity is normally tightly regulated post-translationally and therefore cannot be accurately estimated based on mRNA or protein expression alone. While several types of zymography approaches to estimate protease activity exist, there remains a need for a robust and reliable technique to measure protease activity in biological tissues. We present a novel quantitative ex vivo zymography (QZ) technology based on Probody(®) therapeutics (Pb-Tx), a novel class of protease-activated cancer therapeutics that contain a substrate linker cleavable by tumor-associated proteases. This approach enables the measurement and comparison of protease activity in biological tissues via the detection of Pb-Tx activation. By exploiting substrate specificity and selectivity, cataloguing and differentiating protease activities is possible, with further refinement achieved using protease-specific inhibitors. Using the QZ assay and human tumor xenografts, patient tumor tissues, and patient plasma, we characterized protease activity in preclinical and clinical samples. The QZ assay offers the potential to increase our understanding of protease activity in tissues and inform diagnostic and therapeutic development for diseases, such as cancer, that are characterized by dysregulated proteolysis. MDPI 2021-09-02 /pmc/articles/PMC8471274/ /pubmed/34575469 http://dx.doi.org/10.3390/pharmaceutics13091390 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Howng, Bruce Winter, Michael B. LePage, Carol Popova, Irina Krimm, Michael Vasiljeva, Olga Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies |
title | Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies |
title_full | Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies |
title_fullStr | Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies |
title_full_unstemmed | Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies |
title_short | Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies |
title_sort | novel ex vivo zymography approach for assessment of protease activity in tissues with activatable antibodies |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8471274/ https://www.ncbi.nlm.nih.gov/pubmed/34575469 http://dx.doi.org/10.3390/pharmaceutics13091390 |
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