Site-Specific Native Antibody-Conjugated Microbubbles for Molecular Ultrasound Imaging of Hepatocellular Carcinoma.
1/5 보강
[PURPOSE] To develop a microbubble-based contrast agent using light-activated site-specific conjugation (LASIC) for targeted molecular ultrasound imaging of glypican-3 (GPC3) positive hepatocellular c
APA
Hui J, Shin Y, et al. (2026). Site-Specific Native Antibody-Conjugated Microbubbles for Molecular Ultrasound Imaging of Hepatocellular Carcinoma.. Research square. https://doi.org/10.21203/rs.3.rs-8655411/v1
MLA
Hui J, et al.. "Site-Specific Native Antibody-Conjugated Microbubbles for Molecular Ultrasound Imaging of Hepatocellular Carcinoma.." Research square, 2026.
PMID
41727623
Abstract
[PURPOSE] To develop a microbubble-based contrast agent using light-activated site-specific conjugation (LASIC) for targeted molecular ultrasound imaging of glypican-3 (GPC3) positive hepatocellular carcinoma (HCC).
[MATERIALS AND METHODS] Azide-functionalized microbubbles were conjugated with commercially available anti-GPC3 antibodies using a LASIC DBCO adapter, which enables precise site-specific labeling at the antibody's heavy chain without compromising antigen-binding affinity. 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 seconds, 30 seconds, and 10 minutes post-injection to assess tumor signal enhancement and persistence.
[RESULTS] 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 seconds of injection, persisting up to 10 minutes. In contrast, control microbubbles showed diminished signal intensity post-injection.
[CONCLUSION] 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.
[MATERIALS AND METHODS] Azide-functionalized microbubbles were conjugated with commercially available anti-GPC3 antibodies using a LASIC DBCO adapter, which enables precise site-specific labeling at the antibody's heavy chain without compromising antigen-binding affinity. 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 seconds, 30 seconds, and 10 minutes post-injection to assess tumor signal enhancement and persistence.
[RESULTS] 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 seconds of injection, persisting up to 10 minutes. In contrast, control microbubbles showed diminished signal intensity post-injection.
[CONCLUSION] 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.