Molecular insights into recognition of GUCY2C by T-cell engaging bispecific antibody anti-GUCY2CxCD3
Abstract The intestinal epithelial receptor Guanylyl Cyclase C (GUCY2C) is a tumor-associated cell surface antigen expressed across gastrointestinal malignancies that can serve as an efficacious target for colorectal cancer immunotherapy. Here, we describe a yeast surface-display approach combined w...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-08-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-40467-0 |
| _version_ | 1797452683608588288 |
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| author | Pragya Rampuria Lidia Mosyak Adam R. Root Kristine Svenson Michael J. Agostino Edward R. LaVallie |
| author_facet | Pragya Rampuria Lidia Mosyak Adam R. Root Kristine Svenson Michael J. Agostino Edward R. LaVallie |
| author_sort | Pragya Rampuria |
| collection | DOAJ |
| description | Abstract The intestinal epithelial receptor Guanylyl Cyclase C (GUCY2C) is a tumor-associated cell surface antigen expressed across gastrointestinal malignancies that can serve as an efficacious target for colorectal cancer immunotherapy. Here, we describe a yeast surface-display approach combined with an orthogonal peptide-based mapping strategy to identify the GUCY2C binding epitope of a novel anti-GUCY2CxCD3 bispecific antibody (BsAb) that recently advanced into the clinic for the treatment of cancer. The target epitope was localized to the N-terminal helix H2 of human GUCY2C, which enabled the determination of the crystal structure of the minimal GUCY2C epitope in complex with the anti-GUCY2C antibody domain. To understand if this minimal epitope covers the entire antibody binding region and to investigate the impact of epitope position on the antibody’s activity, we further determined the structure of this interaction in the context of the full-length extracellular domain (ECD) of GUCY2C. We found that this epitope is positioned on the protruding membrane-distal helical region of GUCY2C and that its specific location on the surface of GUCY2C dictates the close spatial proximity of the two antigen arms in a diabody arrangement essential to the tumor killing activity of GUCY2CxCD3 BsAb. |
| first_indexed | 2024-03-09T15:12:12Z |
| format | Article |
| id | doaj.art-db669487a90c46ae8d16d8a5021ff01b |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-09T15:12:12Z |
| publishDate | 2023-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-db669487a90c46ae8d16d8a5021ff01b2023-11-26T13:18:36ZengNature PortfolioScientific Reports2045-23222023-08-0113111510.1038/s41598-023-40467-0Molecular insights into recognition of GUCY2C by T-cell engaging bispecific antibody anti-GUCY2CxCD3Pragya Rampuria0Lidia Mosyak1Adam R. Root2Kristine Svenson3Michael J. Agostino4Edward R. LaVallie5Biomedicine Design, Pfizer Inc.Biomedicine Design, Pfizer Inc.Generate Biomedicines IncBiomedicine Design, Pfizer Inc.Pfizer Digital, Pfizer Inc.Biomedicine Design, Pfizer Inc.Abstract The intestinal epithelial receptor Guanylyl Cyclase C (GUCY2C) is a tumor-associated cell surface antigen expressed across gastrointestinal malignancies that can serve as an efficacious target for colorectal cancer immunotherapy. Here, we describe a yeast surface-display approach combined with an orthogonal peptide-based mapping strategy to identify the GUCY2C binding epitope of a novel anti-GUCY2CxCD3 bispecific antibody (BsAb) that recently advanced into the clinic for the treatment of cancer. The target epitope was localized to the N-terminal helix H2 of human GUCY2C, which enabled the determination of the crystal structure of the minimal GUCY2C epitope in complex with the anti-GUCY2C antibody domain. To understand if this minimal epitope covers the entire antibody binding region and to investigate the impact of epitope position on the antibody’s activity, we further determined the structure of this interaction in the context of the full-length extracellular domain (ECD) of GUCY2C. We found that this epitope is positioned on the protruding membrane-distal helical region of GUCY2C and that its specific location on the surface of GUCY2C dictates the close spatial proximity of the two antigen arms in a diabody arrangement essential to the tumor killing activity of GUCY2CxCD3 BsAb.https://doi.org/10.1038/s41598-023-40467-0 |
| spellingShingle | Pragya Rampuria Lidia Mosyak Adam R. Root Kristine Svenson Michael J. Agostino Edward R. LaVallie Molecular insights into recognition of GUCY2C by T-cell engaging bispecific antibody anti-GUCY2CxCD3 Scientific Reports |
| title | Molecular insights into recognition of GUCY2C by T-cell engaging bispecific antibody anti-GUCY2CxCD3 |
| title_full | Molecular insights into recognition of GUCY2C by T-cell engaging bispecific antibody anti-GUCY2CxCD3 |
| title_fullStr | Molecular insights into recognition of GUCY2C by T-cell engaging bispecific antibody anti-GUCY2CxCD3 |
| title_full_unstemmed | Molecular insights into recognition of GUCY2C by T-cell engaging bispecific antibody anti-GUCY2CxCD3 |
| title_short | Molecular insights into recognition of GUCY2C by T-cell engaging bispecific antibody anti-GUCY2CxCD3 |
| title_sort | molecular insights into recognition of gucy2c by t cell engaging bispecific antibody anti gucy2cxcd3 |
| url | https://doi.org/10.1038/s41598-023-40467-0 |
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