“Stapling” scFv for multispecific biotherapeutics of superior properties
Single-chain fragment variable (scFv) domains play an important role in antibody-based therapeutic modalities, such as bispecifics, multispecifics and chimeric antigen receptor T cells or natural killer cells. However, scFv domains exhibit lower stability and increased risk of aggregation due to tra...
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| Format: | Article |
| 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/10.1080/19420862.2023.2195517 |
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| _version_ | 1849316272987176960 |
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| author | Lauren E. Boucher Elisabeth Geyer Prinslow Michael Feldkamp Fang Yi Rupesh Nanjunda Sheng-Jiun Wu Tun Liu Eilyn R. Lacy Steven Jacobs Natalia Kozlyuk Brian Del Rosario Bingyuan Wu Patricia Aquino Robert C. Davidson Samantha Heyne Nicholas Mazzanti James Testa Michael D. Diem Elsa Gorre Andrew Mahan Hirsh Nanda Harsha P. Gunawardena Alexis Gervais Anthony A. Armstrong Alexey Teplyakov Chichi Huang Adam Zwolak Partha Chowdhury Wan Cheung Cheung Jinquan Luo |
| author_facet | Lauren E. Boucher Elisabeth Geyer Prinslow Michael Feldkamp Fang Yi Rupesh Nanjunda Sheng-Jiun Wu Tun Liu Eilyn R. Lacy Steven Jacobs Natalia Kozlyuk Brian Del Rosario Bingyuan Wu Patricia Aquino Robert C. Davidson Samantha Heyne Nicholas Mazzanti James Testa Michael D. Diem Elsa Gorre Andrew Mahan Hirsh Nanda Harsha P. Gunawardena Alexis Gervais Anthony A. Armstrong Alexey Teplyakov Chichi Huang Adam Zwolak Partha Chowdhury Wan Cheung Cheung Jinquan Luo |
| author_sort | Lauren E. Boucher |
| collection | DOAJ |
| description | Single-chain fragment variable (scFv) domains play an important role in antibody-based therapeutic modalities, such as bispecifics, multispecifics and chimeric antigen receptor T cells or natural killer cells. However, scFv domains exhibit lower stability and increased risk of aggregation due to transient dissociation (“breathing”) and inter-molecular reassociation of the two domains (VL and VH). We designed a novel strategy, referred to as stapling, that introduces two disulfide bonds between the scFv linker and the two variable domains to minimize scFv breathing. We named the resulting molecules stapled scFv (spFv). Stapling increased thermal stability (Tm) by an average of 10°C. In multiple scFv/spFv multispecifics, the spFv molecules display significantly improved stability, minimal aggregation and superior product quality. These spFv multispecifics retain binding affinity and functionality. Our stapling design was compatible with all antibody variable regions we evaluated and may be widely applicable to stabilize scFv molecules for designing biotherapeutics with superior biophysical properties. |
| format | Article |
| id | doaj-art-07a49e5767d741f0bc24b50e3af689e1 |
| institution | Kabale University |
| issn | 1942-0862 1942-0870 |
| language | English |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj-art-07a49e5767d741f0bc24b50e3af689e12025-08-20T03:51:54ZengTaylor & Francis GroupmAbs1942-08621942-08702023-12-0115110.1080/19420862.2023.2195517“Stapling” scFv for multispecific biotherapeutics of superior propertiesLauren E. Boucher0Elisabeth Geyer PrinslowMichael Feldkamp1Fang Yi2Rupesh Nanjunda3Sheng-Jiun Wu4Tun Liu5Eilyn R. Lacy6Steven Jacobs7Natalia Kozlyuk8Brian Del Rosario9Bingyuan Wu10Patricia Aquino11Robert C. Davidson12Samantha Heyne13Nicholas Mazzanti14James Testa15Michael D. Diem16Elsa Gorre17Andrew Mahan18Hirsh Nanda19Harsha P. Gunawardena20Alexis Gervais21Anthony A. Armstrong22Alexey Teplyakov23Chichi Huang24Adam Zwolak25Partha Chowdhury26Wan Cheung Cheung27Jinquan Luo28AbbVie, 1 North Waukegan Rd, North Chicago, IL, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USABiologics Discovery, Janssen Research & Development, LLC, Spring House, PA, USASingle-chain fragment variable (scFv) domains play an important role in antibody-based therapeutic modalities, such as bispecifics, multispecifics and chimeric antigen receptor T cells or natural killer cells. However, scFv domains exhibit lower stability and increased risk of aggregation due to transient dissociation (“breathing”) and inter-molecular reassociation of the two domains (VL and VH). We designed a novel strategy, referred to as stapling, that introduces two disulfide bonds between the scFv linker and the two variable domains to minimize scFv breathing. We named the resulting molecules stapled scFv (spFv). Stapling increased thermal stability (Tm) by an average of 10°C. In multiple scFv/spFv multispecifics, the spFv molecules display significantly improved stability, minimal aggregation and superior product quality. These spFv multispecifics retain binding affinity and functionality. Our stapling design was compatible with all antibody variable regions we evaluated and may be widely applicable to stabilize scFv molecules for designing biotherapeutics with superior biophysical properties.https://www.tandfonline.com/doi/10.1080/19420862.2023.2195517Multi-specific antibodyScfvScfv stabilizationSingle chain fvSpfvStapled scfv |
| spellingShingle | Lauren E. Boucher Elisabeth Geyer Prinslow Michael Feldkamp Fang Yi Rupesh Nanjunda Sheng-Jiun Wu Tun Liu Eilyn R. Lacy Steven Jacobs Natalia Kozlyuk Brian Del Rosario Bingyuan Wu Patricia Aquino Robert C. Davidson Samantha Heyne Nicholas Mazzanti James Testa Michael D. Diem Elsa Gorre Andrew Mahan Hirsh Nanda Harsha P. Gunawardena Alexis Gervais Anthony A. Armstrong Alexey Teplyakov Chichi Huang Adam Zwolak Partha Chowdhury Wan Cheung Cheung Jinquan Luo “Stapling” scFv for multispecific biotherapeutics of superior properties mAbs Multi-specific antibody Scfv Scfv stabilization Single chain fv Spfv Stapled scfv |
| title | “Stapling” scFv for multispecific biotherapeutics of superior properties |
| title_full | “Stapling” scFv for multispecific biotherapeutics of superior properties |
| title_fullStr | “Stapling” scFv for multispecific biotherapeutics of superior properties |
| title_full_unstemmed | “Stapling” scFv for multispecific biotherapeutics of superior properties |
| title_short | “Stapling” scFv for multispecific biotherapeutics of superior properties |
| title_sort | stapling scfv for multispecific biotherapeutics of superior properties |
| topic | Multi-specific antibody Scfv Scfv stabilization Single chain fv Spfv Stapled scfv |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2023.2195517 |
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