Comparative Characterization of Different Molecular Formats of Bispecific Antibodies Targeting EGFR and PD-L1
We generated two IgG1-like bispecific antibodies (BsAbs) with different molecular formats, symmetrical DVD-Ig and asymmetrical knob-in-hole (KIH), targeting the same antigens, EGFR and PD-L1 (designated as anti-EGFR/PD-L1). We performed the physiochemical and biological characterization of these two...
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MDPI AG
2022-06-01
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| Online Access: | https://www.mdpi.com/1999-4923/14/7/1381 |
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| author | Nishant Mohan Atul Agrawal Yi Shen Katie L. Winarski Yukinori Endo Milos Dokmanovic Deborah Schmiel Jiwen Zheng David S. Rotstein Lorraine C. Pelosof Wen Jin Wu |
| author_facet | Nishant Mohan Atul Agrawal Yi Shen Katie L. Winarski Yukinori Endo Milos Dokmanovic Deborah Schmiel Jiwen Zheng David S. Rotstein Lorraine C. Pelosof Wen Jin Wu |
| author_sort | Nishant Mohan |
| collection | DOAJ |
| description | We generated two IgG1-like bispecific antibodies (BsAbs) with different molecular formats, symmetrical DVD-Ig and asymmetrical knob-in-hole (KIH), targeting the same antigens, EGFR and PD-L1 (designated as anti-EGFR/PD-L1). We performed the physiochemical and biological characterization of these two formats of anti-EGFR/PD-L1 BsAbs and compared some key quality attributes and biological activities of these two formats of BsAbs. Physiochemical binding characterization data demonstrated that both formats bound EGFR and PD-L1. However, the binding affinity of the KIH format was weaker than the DVD-Ig format in Biacore binding assays. In contrast, both DVD-Ig and KIH BsAbs had similar ELISA and cell surface binding activities, comparable to mAbs. Triple-negative breast cancer (TNBC) cells and a xenograft model were used to test the potency of BsAbs and other biological activities. Results showed that anti-EGFR/PD-L1 BsAbs exhibited in vitro and in vivo antitumor proliferation activity, but there was a difference in the potencies of the respective BsAb formats (DVD-Ig and KIH) when different cells or assays were used. This study provides evidence that the potency of the BsAbs targeting the same antigens can be affected by the respective molecular features, and selection of appropriate cell lines and assays is critically important for the assay development and potency testing of BsAbs. |
| first_indexed | 2024-03-09T06:04:17Z |
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| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T06:04:17Z |
| publishDate | 2022-06-01 |
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| series | Pharmaceutics |
| spelling | doaj.art-58be08b2889c41238320db318c0f3eb42023-12-03T12:06:08ZengMDPI AGPharmaceutics1999-49232022-06-01147138110.3390/pharmaceutics14071381Comparative Characterization of Different Molecular Formats of Bispecific Antibodies Targeting EGFR and PD-L1Nishant Mohan0Atul Agrawal1Yi Shen2Katie L. Winarski3Yukinori Endo4Milos Dokmanovic5Deborah Schmiel6Jiwen Zheng7David S. Rotstein8Lorraine C. Pelosof9Wen Jin Wu10Division of Biotechnology Review and Research 1, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Biotechnology Review and Research 1, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Biotechnology Review and Research 1, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Biotechnology Review and Research 1, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Biotechnology Review and Research 1, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Biotechnology Review and Research 1, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Biotechnology Review and Research 1, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Biology, Chemistry and Materials Science, Office of Science and Engineering Laboratories, Center for Devices and Radiological Health, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Compliance, Office of Surveillance and Compliance, Center for Veterinary Medicine, U.S. Food and Drug Administration, Derwood, MD 20855, USADivision of Oncology 3, Office of Oncologic Diseases, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USADivision of Biotechnology Review and Research 1, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD 20993, USAWe generated two IgG1-like bispecific antibodies (BsAbs) with different molecular formats, symmetrical DVD-Ig and asymmetrical knob-in-hole (KIH), targeting the same antigens, EGFR and PD-L1 (designated as anti-EGFR/PD-L1). We performed the physiochemical and biological characterization of these two formats of anti-EGFR/PD-L1 BsAbs and compared some key quality attributes and biological activities of these two formats of BsAbs. Physiochemical binding characterization data demonstrated that both formats bound EGFR and PD-L1. However, the binding affinity of the KIH format was weaker than the DVD-Ig format in Biacore binding assays. In contrast, both DVD-Ig and KIH BsAbs had similar ELISA and cell surface binding activities, comparable to mAbs. Triple-negative breast cancer (TNBC) cells and a xenograft model were used to test the potency of BsAbs and other biological activities. Results showed that anti-EGFR/PD-L1 BsAbs exhibited in vitro and in vivo antitumor proliferation activity, but there was a difference in the potencies of the respective BsAb formats (DVD-Ig and KIH) when different cells or assays were used. This study provides evidence that the potency of the BsAbs targeting the same antigens can be affected by the respective molecular features, and selection of appropriate cell lines and assays is critically important for the assay development and potency testing of BsAbs.https://www.mdpi.com/1999-4923/14/7/1381bispecific antibodies (BsAbs)EFGRPD-L1triple-negative breast cancer (TNBC)bioassays |
| spellingShingle | Nishant Mohan Atul Agrawal Yi Shen Katie L. Winarski Yukinori Endo Milos Dokmanovic Deborah Schmiel Jiwen Zheng David S. Rotstein Lorraine C. Pelosof Wen Jin Wu Comparative Characterization of Different Molecular Formats of Bispecific Antibodies Targeting EGFR and PD-L1 Pharmaceutics bispecific antibodies (BsAbs) EFGR PD-L1 triple-negative breast cancer (TNBC) bioassays |
| title | Comparative Characterization of Different Molecular Formats of Bispecific Antibodies Targeting EGFR and PD-L1 |
| title_full | Comparative Characterization of Different Molecular Formats of Bispecific Antibodies Targeting EGFR and PD-L1 |
| title_fullStr | Comparative Characterization of Different Molecular Formats of Bispecific Antibodies Targeting EGFR and PD-L1 |
| title_full_unstemmed | Comparative Characterization of Different Molecular Formats of Bispecific Antibodies Targeting EGFR and PD-L1 |
| title_short | Comparative Characterization of Different Molecular Formats of Bispecific Antibodies Targeting EGFR and PD-L1 |
| title_sort | comparative characterization of different molecular formats of bispecific antibodies targeting egfr and pd l1 |
| topic | bispecific antibodies (BsAbs) EFGR PD-L1 triple-negative breast cancer (TNBC) bioassays |
| url | https://www.mdpi.com/1999-4923/14/7/1381 |
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