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Here, we develop a single-cell plasmonic imaging platform to quantitatively resolve the interaction between the membrane human epidermal growth factor receptor 2 (HER2) and HER2-targeted therapeutics. This label-free approach enables continuous monitoring of the molecular interaction process, allowing real-time extraction of detailed binding kinetics. Using this platform, we systematically compare the binding behaviors of the HER2-targeting antibody trastuzumab (Herceptin) and clinically relevant ADCs (T-DM1 and T-DXd), revealing distinct kinetic signatures associated with the drug conjugation. Analysis across cell lines with different HER2 expressions reveals that increased receptor density does not necessarily enhance binding stability, suggesting a potential trade-off between receptor availability and effective interaction dynamics. To further evaluate the potential capability of tracking membrane-associated dynamics, polystyrene nanoparticle probes were employed to validate real-time imaging of endocytosis dynamics, distinguishing uptake behavior in live versus fixed cells.This work establishes cell-based plasmonic imaging as a quantitative and mechanistic approach for evaluating ADC-receptor interactions in situ, offering valuable insights for rational ADC design and precision therapeutic optimization.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c02515","type":"journal-article","created":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T08:01:53Z","timestamp":1786608113000},"page":"24226-24236","source":"Crossref","is-referenced-by-count":0,"title":["<i>In Situ<\/i>\n                    Tracking of Membrane HER2\n- Antibody\u2013Drug Conjugate Interaction with Surface Plasmon\nResonance Imaging"],"prefix":"10.1021","volume":"98","author":[{"given":"Haiying","family":"Ding","sequence":"first","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, P. R. China, 310058"]},{"name":"Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences , , ,","place":["Hangzhou, P. R. China, 310005"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyin","family":"Liu","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, P. R. China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingxue","family":"Guo","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, P. R. China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yueping","family":"Qiu","sequence":"additional","affiliation":[{"name":"Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences , , ,","place":["Hangzhou, P. R. China, 310005"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyu","family":"Wu","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, P. R. China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunxiao","family":"Wang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, P. R. China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baiqi","family":"Cui","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, P. R. China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luo","family":"Fang","sequence":"additional","affiliation":[{"name":"Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences , , ,","place":["Hangzhou, P. R. China, 310005"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4369-8563","authenticated-orcid":true,"given":"Fenni","family":"Zhang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, P. R. 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