Generation and engineering of potent single domain antibody-based bispecific IL-18 mimetics resistant to IL-18BP decoy receptor inhibition
Here, we generated bispecific antibody (bsAb) derivatives that mimic the function of interleukin (IL)-18 based on single domain antibodies (sdAbs) specific to IL-18 Rα and IL-18 Rβ. For this, camelids were immunized, followed by yeast surface display (YSD)-enabled discovery of VHHs targeting the ind...
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| Main Authors: | , , , , , , , , , , , , , , , , |
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| Language: | English |
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Taylor & Francis Group
2023-12-01
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| Series: | mAbs |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2023.2236265 |
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| author | Britta Lipinski Laura Unmuth Paul Arras Stefan Becker Christina Bauer Lars Toleikis Simon Krah Achim Doerner Desislava Yanakieva Ammelie Svea Boje Katja Klausz Matthias Peipp Vanessa Siegmund Andreas Evers Harald Kolmar Lukas Pekar Stefan Zielonka |
| author_facet | Britta Lipinski Laura Unmuth Paul Arras Stefan Becker Christina Bauer Lars Toleikis Simon Krah Achim Doerner Desislava Yanakieva Ammelie Svea Boje Katja Klausz Matthias Peipp Vanessa Siegmund Andreas Evers Harald Kolmar Lukas Pekar Stefan Zielonka |
| author_sort | Britta Lipinski |
| collection | DOAJ |
| description | Here, we generated bispecific antibody (bsAb) derivatives that mimic the function of interleukin (IL)-18 based on single domain antibodies (sdAbs) specific to IL-18 Rα and IL-18 Rβ. For this, camelids were immunized, followed by yeast surface display (YSD)-enabled discovery of VHHs targeting the individual receptor subunits. Upon reformatting into a strictly monovalent (1 + 1) bispecific sdAb architecture, several bsAbs triggered dose-dependent IL-18 R downstream signaling on IL-18 reporter cells, as well as IFN-γ release by peripheral blood mononuclear cells in the presence of low-dose IL-12. However, compared with IL-18, potencies and efficacies were considerably attenuated. By engineering paratope valencies and the spatial orientation of individual paratopes within the overall design architecture, we were able to generate IL-18 mimetics displaying significantly augmented functionalities, resulting in bispecific cytokine mimetics that were more potent than IL-18 in triggering proinflammatory cytokine release. Furthermore, generated IL-18 mimetics were unaffected from inhibition by IL-18 binding protein decoy receptor. Essentially, we demonstrate that this strategy enables the generation of IL-18 mimetics with tailor-made cytokine functionalities. |
| format | Article |
| id | doaj-art-7e0581bda58843b2a64abbd81a6ee1ef |
| institution | DOAJ |
| issn | 1942-0862 1942-0870 |
| language | English |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj-art-7e0581bda58843b2a64abbd81a6ee1ef2025-08-20T02:47:49ZengTaylor & Francis GroupmAbs1942-08621942-08702023-12-0115110.1080/19420862.2023.2236265Generation and engineering of potent single domain antibody-based bispecific IL-18 mimetics resistant to IL-18BP decoy receptor inhibitionBritta Lipinski0Laura Unmuth1Paul Arras2Stefan Becker3Christina Bauer4Lars Toleikis5Simon Krah6Achim Doerner7Desislava Yanakieva8Ammelie Svea Boje9Katja Klausz10Matthias Peipp11Vanessa Siegmund12Andreas Evers13Harald Kolmar14Lukas Pekar15Stefan Zielonka16Antibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyEarly Protein Supply and Characterization (EPSC), Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyEarly Protein Supply and Characterization (EPSC), Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyEarly Protein Supply and Characterization (EPSC), Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyDivision of Antibody-Based Immunotherapy, Department of Internal Medicine II, University Hospital Schleswig-Holstein and Christian-Albrechts-University Kiel, Kiel, GermanyDivision of Antibody-Based Immunotherapy, Department of Internal Medicine II, University Hospital Schleswig-Holstein and Christian-Albrechts-University Kiel, Kiel, GermanyDivision of Antibody-Based Immunotherapy, Department of Internal Medicine II, University Hospital Schleswig-Holstein and Christian-Albrechts-University Kiel, Kiel, GermanyEarly Protein Supply and Characterization (EPSC), Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyInstitute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, GermanyAntibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyAntibody Discovery and Protein Engineering (ADPE), Merck Healthcare KGaA, Darmstadt, GermanyHere, we generated bispecific antibody (bsAb) derivatives that mimic the function of interleukin (IL)-18 based on single domain antibodies (sdAbs) specific to IL-18 Rα and IL-18 Rβ. For this, camelids were immunized, followed by yeast surface display (YSD)-enabled discovery of VHHs targeting the individual receptor subunits. Upon reformatting into a strictly monovalent (1 + 1) bispecific sdAb architecture, several bsAbs triggered dose-dependent IL-18 R downstream signaling on IL-18 reporter cells, as well as IFN-γ release by peripheral blood mononuclear cells in the presence of low-dose IL-12. However, compared with IL-18, potencies and efficacies were considerably attenuated. By engineering paratope valencies and the spatial orientation of individual paratopes within the overall design architecture, we were able to generate IL-18 mimetics displaying significantly augmented functionalities, resulting in bispecific cytokine mimetics that were more potent than IL-18 in triggering proinflammatory cytokine release. Furthermore, generated IL-18 mimetics were unaffected from inhibition by IL-18 binding protein decoy receptor. Essentially, we demonstrate that this strategy enables the generation of IL-18 mimetics with tailor-made cytokine functionalities.https://www.tandfonline.com/doi/10.1080/19420862.2023.2236265Antibody engineeringbispecific antibodycytokine mimeticIL-18IL-18 binding proteinIL-18BP |
| spellingShingle | Britta Lipinski Laura Unmuth Paul Arras Stefan Becker Christina Bauer Lars Toleikis Simon Krah Achim Doerner Desislava Yanakieva Ammelie Svea Boje Katja Klausz Matthias Peipp Vanessa Siegmund Andreas Evers Harald Kolmar Lukas Pekar Stefan Zielonka Generation and engineering of potent single domain antibody-based bispecific IL-18 mimetics resistant to IL-18BP decoy receptor inhibition mAbs Antibody engineering bispecific antibody cytokine mimetic IL-18 IL-18 binding protein IL-18BP |
| title | Generation and engineering of potent single domain antibody-based bispecific IL-18 mimetics resistant to IL-18BP decoy receptor inhibition |
| title_full | Generation and engineering of potent single domain antibody-based bispecific IL-18 mimetics resistant to IL-18BP decoy receptor inhibition |
| title_fullStr | Generation and engineering of potent single domain antibody-based bispecific IL-18 mimetics resistant to IL-18BP decoy receptor inhibition |
| title_full_unstemmed | Generation and engineering of potent single domain antibody-based bispecific IL-18 mimetics resistant to IL-18BP decoy receptor inhibition |
| title_short | Generation and engineering of potent single domain antibody-based bispecific IL-18 mimetics resistant to IL-18BP decoy receptor inhibition |
| title_sort | generation and engineering of potent single domain antibody based bispecific il 18 mimetics resistant to il 18bp decoy receptor inhibition |
| topic | Antibody engineering bispecific antibody cytokine mimetic IL-18 IL-18 binding protein IL-18BP |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2023.2236265 |
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