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Binding specificity was confirmed through pre-blocking experiments with free anti-GPC3 antibodies in HCC cell lines. Fluorescence microscopy and quantitative image analysis were employed to evaluate cell-binding efficiency. For in vivo validation, conjugated microbubbles were intravenously administered into mice bearing orthotopic HepG2 xenografts, followed by contrast-enhanced ultrasound (CEUS) imaging at 0\u00a0s, 30\u00a0s, and 10\u00a0min post-injection to assess tumor signal enhancement and persistence.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>LASIC chemistry enabled covalent and site-specific antibody attachment to microbubbles. In vitro assays demonstrated significantly increased binding of targeted microbubbles to GPC3-positive cells compared to antigen-blocked controls. In vivo CEUS imaging revealed rapid and sustained tumor-specific signal enhancement within 30\u00a0s of injection, persisting up to 10\u00a0min. In contrast, control microbubbles showed diminished signal intensity\u00a0and rapid wash out post-injection.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>LASIC-conjugated anti-GPC3 microbubbles exhibit efficient, specific, and durable binding to GPC3-expressing HCC cells both in vitro and in vivo. This strategy enables real-time, image-guided molecular profiling via CEUS and holds promise for precision imaging and theranostic applications in liver cancer management.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s11307-026-02131-7","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T20:06:12Z","timestamp":1786478772000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Site-Specific Native Antibody-Conjugated Microbubbles for Molecular Ultrasound Imaging of Hepatocellular Carcinoma"],"prefix":"10.1007","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0876-9458","authenticated-orcid":false,"given":"James","family":"Hui","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yonghwan","family":"Shin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janet","family":"Pham","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Po-Chun","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aravinth","family":"Ruppa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veronica","family":"Hankil","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lea","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanif Saifurrahman","family":"Kalamullah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naoki","family":"Kaneko","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2896-1930","authenticated-orcid":false,"given":"Jason","family":"Chiang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,11]]},"reference":[{"issue":"1","key":"2131_CR1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-020-00240-3","volume":"7","author":"JM Llovet","year":"2021","unstructured":"Llovet JM, Kelley RK, Villanueva A, Singal AG, Pikarsky E, Roayaie S (2021) Hepatocellular carcinoma. 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