Modular Conjugation of a Potent Anti-HER2 Immunotoxin Using Coassociating Peptides
| العنوان: | Modular Conjugation of a Potent Anti-HER2 Immunotoxin Using Coassociating Peptides |
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| المؤلفون: | Mariel Donzeau, Celia Deville, Yves Nominé, Nikita Pallaoro, Lina Barret, Bruno Chatton, Marie Blocat, Ambre Bender, Audrey Stoessel, Lucile Guyot, Leonel Nguekeu-Zebaze, Guy Zuber, Nadja Groysbeck, Laetitia Voilquin, Murielle Masson, Thomas Lutz |
| المساهمون: | Biotechnologie et signalisation cellulaire (BSC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut de recherche de l'Ecole de biotechnologie de Strasbourg (IREBS), Centre for Integrative Biology - CBI (Inserm U964 - CNRS UMR7104 - IGBMC), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS) |
| المصدر: | Bioconjugate Chemistry Bioconjugate Chemistry, American Chemical Society, 2020, 31 (10), pp.2421-2430. ⟨10.1021/acs.bioconjchem.0c00482⟩ |
| بيانات النشر: | HAL CCSD, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Models, Molecular, Receptor, ErbB-2, Virulence Factors, Recombinant Fusion Proteins, Bacterial Toxins, Biomedical Engineering, Pharmaceutical Science, Exotoxins, Bioengineering, Antineoplastic Agents, Breast Neoplasms, medicine.disease_cause, Immunotoxin, Cell Line, Tumor, medicine, Humans, Cytotoxicity, skin and connective tissue diseases, Escherichia coli, Binding selectivity, Pharmacology, ADP Ribose Transferases, Sciences du Vivant [q-bio]/Ingénierie biomédicale, Toxin, Chemistry, Immunotoxins, Organic Chemistry, Single-Domain Antibodies, Fusion protein, 3. Good health, Biochemistry, Covalent bond, bacteria, Female, [SDV.IB]Life Sciences [q-bio]/Bioengineering, Biotechnology, Conjugate |
| الوصف: | Immunotoxins are emerging candidates for cancer therapeutics. These biomolecules consist of a cell-targeting protein combined to a polypeptide toxin. Associations of both entities can be achieved either chemically by covalent bonds or genetically creating fusion proteins. However, chemical agents can affect the activity and/or stability of the conjugate proteins, and additional purification steps are often required to isolate the final conjugate from unwanted byproducts. As for fusion proteins, they often suffer from low solubility and yield. In this report, we describe a straightforward conjugation process to generate an immunotoxin using coassociating peptides (named K3 and E3), originating from the tetramerization domain of p53. To that end, a nanobody targeting the human epidermal growth factor receptor 2 (nano-HER2) and a protein toxin fragment from Pseudomonas aeruginosa exotoxin A (TOX) were genetically fused to the E3 and K3 peptides. Entities were produced separately in Escherichia coli in soluble forms and at high yields. The nano-HER2 fused to the E3 or K3 helixes (nano-HER2-E3 and nano-HER2-K3) and the coassembled immunotoxins (nano-HER2-K3E3-TOX and nano-HER2-E3K3-TOX) presented binding specificity on HER2-overexpressing cells with relative binding constants in the low nanomolar to picomolar range. Both toxin modules (E3-TOX and K3-TOX) and the combined immunotoxins exhibited similar cytotoxicity levels compared to the toxin alone (TOX). Finally, nano-HER2-K3E3-TOX and nano-HER2-E3K3-TOX evaluated on various breast cancer cells were highly potent and specific to killing HER2-overexpressing breast cancer cells with IC50 values in the picomolar range. Altogether, we demonstrate that this noncovalent conjugation method using two coassembling peptides can be easily implemented for the modular engineering of immunotoxins targeting different types of cancers. |
| اللغة: | English |
| تدمد: | 1043-1802 1520-4812 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c0f2c9a574f7317a827404a0d150ea01Test https://hal.archives-ouvertes.fr/hal-03005245Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....c0f2c9a574f7317a827404a0d150ea01 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10431802 15204812 |
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