Shortwave-infrared (SWIR) fluorescence molecular imaging using indocyanine green–antibody conjugates for the optical diagnostics of cancerous tumours

Abstract

Recently, shortwave-infrared (SWIR, 1000–1400 nm) fluorescence imaging has attracted much attention due to the higher contrast and sensitivity with deeper penetration depths compared to conventional visible and near-infrared (NIR) fluorescence imaging. For the SWIR fluorescence imaging, the development of fluorescent probes emitting over 1000 nm is necessary. So far, a variety of SWIR fluorescent probes based on single-walled carbon nanotubes, quantum dots, rare-metal doped nanomaterials, and organic dyes have been developed. However, there are a very limited number of biocompatible SWIR fluorescent probes, which can be used to biomedical applications. Among NIR and SWIR fluorescent probes, indocyanine green (ICG) is the only fluorescent dye approved by US Food and Drug Administration (FDA) for clinical use. Although ICG has a fluorescence maximum at a NIR region (ca. 830 nm), ICG emits in the SWIR region over 1000 nm. Here, we present ICG-based SWIR fluorescence molecular imaging for the highly-sensitive optical detection of breast and skin tumours in mice. As SWIR fluorescent molecular-imaging probes, we synthesized ICG–antibody conjugates, which prepared from anti-HER2 antibody (Herceptin), anti-EGFR antibody (Erbitux), anti-VEGFR-2 antibody (Cyramza), and anti-PD-L1 antibody (anti-PD-L1 ab). The present SWIR molecular imaging probes specifically accumulated to the breast and skin tumours, and their SWIR fluorescence images (>1000 nm) showed 1.5–2.0 times higher contrast than NIR tumour images taken at 830 nm. We show that the SWIR fluorescence imaging using ICG–antibody conjugates can be used for the elucidation of expression level of cancer-specific membrane proteins, HER2, EGFR, VEGFR-2, and PD-L1 in vivo. We also show that the SWIR fluorescence imaging enables quantitative analysis of the change in the size of tumour treated with an anti-cancer drug, Kadcyla. Our findings suggest that the SWIR fluorescence molecular imaging using ICG–antibody conjugates has potential to use for the optical diagnostics of cancerous tumors in medical and clinical fields.