Engineering hydrophobicity and manufacturability for optimized biparatopic antibody–drug conjugates targeting c-MET
Optimal combinations of paratopes assembled into a biparatopic antibody have the capacity to mediate high-grade target cross-linking on cell membranes, leading to degradation of the target, as well as antibody and payload delivery in the case of an antibody-drug conjugate (ADC). In the work presente...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2024-12-01
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| Series: | mAbs |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2024.2302386 |
| _version_ | 1826846372610965504 |
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| author | Andreas Evers Simon Krah Deniz Demir Ramona Gaa Desislava Elter Christian Schroeter Stefan Zielonka Nicolas Rasche Julia Dotterweich Christine Knuehl Achim Doerner |
| author_facet | Andreas Evers Simon Krah Deniz Demir Ramona Gaa Desislava Elter Christian Schroeter Stefan Zielonka Nicolas Rasche Julia Dotterweich Christine Knuehl Achim Doerner |
| author_sort | Andreas Evers |
| collection | DOAJ |
| description | Optimal combinations of paratopes assembled into a biparatopic antibody have the capacity to mediate high-grade target cross-linking on cell membranes, leading to degradation of the target, as well as antibody and payload delivery in the case of an antibody-drug conjugate (ADC). In the work presented here, molecular docking suggested a suitable paratope combination targeting c-MET, but hydrophobic patches in essential binding regions of one moiety necessitated engineering. In addition to rational design of HCDR2 and HCDR3 mutations, site-specific spiking libraries were generated and screened in yeast and mammalian surface display approaches. Comparative analyses revealed similar positions amendable for hydrophobicity reduction, with a broad combinatorial diversity obtained from library outputs. Optimized variants showed high stability, strongly reduced hydrophobicity, retained affinities supporting the desired functionality and enhanced producibility. The resulting biparatopic anti-c-MET ADCs were comparably active on c-MET expressing tumor cell lines as REGN5093 exatecan DAR6 ADC. Structural molecular modeling of paratope combinations for preferential inter-target binding combined with protein engineering for manufacturability yielded deep insights into the capabilities of rational and library approaches. The methodologies of in silico hydrophobicity identification and sequence optimization could serve as a blueprint for rapid development of optimal biparatopic ADCs targeting further tumor-associated antigens in the future. |
| first_indexed | 2024-03-08T14:29:21Z |
| format | Article |
| id | doaj.art-ba6e0960fee8491f977fc74b355ed448 |
| institution | Directory Open Access Journal |
| issn | 1942-0862 1942-0870 |
| language | English |
| last_indexed | 2025-02-16T15:00:12Z |
| publishDate | 2024-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj.art-ba6e0960fee8491f977fc74b355ed4482025-01-31T04:19:37ZengTaylor & Francis GroupmAbs1942-08621942-08702024-12-0116110.1080/19420862.2024.2302386Engineering hydrophobicity and manufacturability for optimized biparatopic antibody–drug conjugates targeting c-METAndreas Evers0Simon Krah1Deniz Demir2Ramona Gaa3Desislava Elter4Christian Schroeter5Stefan Zielonka6Nicolas Rasche7Julia Dotterweich8Christine Knuehl9Achim Doerner10Antibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, GermanyADC and Targeted Therapeutics, Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, GermanyADC and Targeted Therapeutics, Merck Healthcare KGaA, Darmstadt, GermanyResearch Unit Oncology, Merck Healthcare KGaA, Darmstadt, GermanyResearch Unit Oncology, Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering, Merck Healthcare KGaA, Darmstadt, GermanyOptimal combinations of paratopes assembled into a biparatopic antibody have the capacity to mediate high-grade target cross-linking on cell membranes, leading to degradation of the target, as well as antibody and payload delivery in the case of an antibody-drug conjugate (ADC). In the work presented here, molecular docking suggested a suitable paratope combination targeting c-MET, but hydrophobic patches in essential binding regions of one moiety necessitated engineering. In addition to rational design of HCDR2 and HCDR3 mutations, site-specific spiking libraries were generated and screened in yeast and mammalian surface display approaches. Comparative analyses revealed similar positions amendable for hydrophobicity reduction, with a broad combinatorial diversity obtained from library outputs. Optimized variants showed high stability, strongly reduced hydrophobicity, retained affinities supporting the desired functionality and enhanced producibility. The resulting biparatopic anti-c-MET ADCs were comparably active on c-MET expressing tumor cell lines as REGN5093 exatecan DAR6 ADC. Structural molecular modeling of paratope combinations for preferential inter-target binding combined with protein engineering for manufacturability yielded deep insights into the capabilities of rational and library approaches. The methodologies of in silico hydrophobicity identification and sequence optimization could serve as a blueprint for rapid development of optimal biparatopic ADCs targeting further tumor-associated antigens in the future.https://www.tandfonline.com/doi/10.1080/19420862.2024.2302386Antibody engineeringBiparatopic ADCc-METhydrophobicitymanufacturability |
| spellingShingle | Andreas Evers Simon Krah Deniz Demir Ramona Gaa Desislava Elter Christian Schroeter Stefan Zielonka Nicolas Rasche Julia Dotterweich Christine Knuehl Achim Doerner Engineering hydrophobicity and manufacturability for optimized biparatopic antibody–drug conjugates targeting c-MET mAbs Antibody engineering Biparatopic ADC c-MET hydrophobicity manufacturability |
| title | Engineering hydrophobicity and manufacturability for optimized biparatopic antibody–drug conjugates targeting c-MET |
| title_full | Engineering hydrophobicity and manufacturability for optimized biparatopic antibody–drug conjugates targeting c-MET |
| title_fullStr | Engineering hydrophobicity and manufacturability for optimized biparatopic antibody–drug conjugates targeting c-MET |
| title_full_unstemmed | Engineering hydrophobicity and manufacturability for optimized biparatopic antibody–drug conjugates targeting c-MET |
| title_short | Engineering hydrophobicity and manufacturability for optimized biparatopic antibody–drug conjugates targeting c-MET |
| title_sort | engineering hydrophobicity and manufacturability for optimized biparatopic antibody drug conjugates targeting c met |
| topic | Antibody engineering Biparatopic ADC c-MET hydrophobicity manufacturability |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2024.2302386 |
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