Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor Targeting
Trophoblast cell-surface antigen 2 (Trop-2) is a tumor-associated antigen that is connected with the development of various tumors and has been identified as a promising target for tumor immunotherapy. To date, the immunotherapy against Trop-2 mainly relies on the specific targeting by monoclonal an...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-07-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/14/7942 |
| _version_ | 1797406083223912448 |
|---|---|
| author | Yaozhong Hu Yi Wang Jing Lin Sihao Wu Huan Lv Xuemeng Ji Shuo Wang |
| author_facet | Yaozhong Hu Yi Wang Jing Lin Sihao Wu Huan Lv Xuemeng Ji Shuo Wang |
| author_sort | Yaozhong Hu |
| collection | DOAJ |
| description | Trophoblast cell-surface antigen 2 (Trop-2) is a tumor-associated antigen that is connected with the development of various tumors and has been identified as a promising target for tumor immunotherapy. To date, the immunotherapy against Trop-2 mainly relies on the specific targeting by monoclonal antibody in antibody-drug conjugate (ADC). Alternatively, the single domain antibodies of nanobodies (Nbs) possesses unique properties such as smaller size, better tissue penetration, etc., to make them good candidates for tumor targeting. Thus, it was proposed to develop anti-Trop-2 Nbs for tumor targeting in this study. Generally, three consecutive rounds of bio-panning were performed against immobilized recombinant Trop-2, and yielded three Nbs (Nb60, Nb65, and Nb108). The affinity of selected Nbs was determined in the nanomolar range, especially the good properties of Nb60 were verified as a promising candidate for tumor labeling. The binding to native Trop-2 was confirmed by flow cytometry against tumor cells. The inhibitory effects of the selected Nbs on tumor cell proliferation and migration were confirmed by wound healing and Transwell assay. The clear localization of the selected Nbs on the surface of tumor cells verified the potent labeling efficiency. In conclusion, this study provided several Nbs with the potential to be developed as targeting moiety of drug conjugates. |
| first_indexed | 2024-03-09T03:20:03Z |
| format | Article |
| id | doaj.art-18a68f5989fc40579392c549e97a3e55 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T03:20:03Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-18a68f5989fc40579392c549e97a3e552023-12-03T15:10:23ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-07-012314794210.3390/ijms23147942Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor TargetingYaozhong Hu0Yi Wang1Jing Lin2Sihao Wu3Huan Lv4Xuemeng Ji5Shuo Wang6Research Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaTrophoblast cell-surface antigen 2 (Trop-2) is a tumor-associated antigen that is connected with the development of various tumors and has been identified as a promising target for tumor immunotherapy. To date, the immunotherapy against Trop-2 mainly relies on the specific targeting by monoclonal antibody in antibody-drug conjugate (ADC). Alternatively, the single domain antibodies of nanobodies (Nbs) possesses unique properties such as smaller size, better tissue penetration, etc., to make them good candidates for tumor targeting. Thus, it was proposed to develop anti-Trop-2 Nbs for tumor targeting in this study. Generally, three consecutive rounds of bio-panning were performed against immobilized recombinant Trop-2, and yielded three Nbs (Nb60, Nb65, and Nb108). The affinity of selected Nbs was determined in the nanomolar range, especially the good properties of Nb60 were verified as a promising candidate for tumor labeling. The binding to native Trop-2 was confirmed by flow cytometry against tumor cells. The inhibitory effects of the selected Nbs on tumor cell proliferation and migration were confirmed by wound healing and Transwell assay. The clear localization of the selected Nbs on the surface of tumor cells verified the potent labeling efficiency. In conclusion, this study provided several Nbs with the potential to be developed as targeting moiety of drug conjugates.https://www.mdpi.com/1422-0067/23/14/7942Trop-2nanobodytumor targetingproliferationmigration |
| spellingShingle | Yaozhong Hu Yi Wang Jing Lin Sihao Wu Huan Lv Xuemeng Ji Shuo Wang Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor Targeting International Journal of Molecular Sciences Trop-2 nanobody tumor targeting proliferation migration |
| title | Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor Targeting |
| title_full | Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor Targeting |
| title_fullStr | Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor Targeting |
| title_full_unstemmed | Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor Targeting |
| title_short | Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor Targeting |
| title_sort | identification and characterization of specific nanobodies against trop 2 for tumor targeting |
| topic | Trop-2 nanobody tumor targeting proliferation migration |
| url | https://www.mdpi.com/1422-0067/23/14/7942 |
| work_keys_str_mv | AT yaozhonghu identificationandcharacterizationofspecificnanobodiesagainsttrop2fortumortargeting AT yiwang identificationandcharacterizationofspecificnanobodiesagainsttrop2fortumortargeting AT jinglin identificationandcharacterizationofspecificnanobodiesagainsttrop2fortumortargeting AT sihaowu identificationandcharacterizationofspecificnanobodiesagainsttrop2fortumortargeting AT huanlv identificationandcharacterizationofspecificnanobodiesagainsttrop2fortumortargeting AT xuemengji identificationandcharacterizationofspecificnanobodiesagainsttrop2fortumortargeting AT shuowang identificationandcharacterizationofspecificnanobodiesagainsttrop2fortumortargeting |