RIKEN Center for Biosystems Dynamics Research (BDR), RIKEN, Furuedai 6-2-3, Suita, Osaka, 565-0874, Japan.
Graduate School of Frontier BioSciences, Osaka University, Yamada-oka 1-3, Suita, Osaka, 565-0871, Japan.
Chembiochem. 2019 Feb 15;20(4):568-575. doi: 10.1002/cbic.201800506. Epub 2018 Dec 5.
Near-infrared (NIR)-emitting fluorescent probes are widely used for molecular imaging at the whole-body level. However, NIR-emitting fluorescent probes emitting over λ=700 nm are not suitable for molecular imaging at the cellular level, because most of the conventional fluorescence microscopes have very low optical sensitivity in the NIR region. Thus, to achieve fluorescence imaging at the cellular and whole-body levels by using single probes, visible and NIR-emitting dual-color fluorescent probes are desirable. For dual-color fluorescence molecular imaging, we synthesized fluorescent, recombinant-protein-conjugated, NIR-emitting quantum dots (QDs), in which the recombinant protein consists of enhanced green fluorescent protein (EGFP) and the immunoglobulin binding domain (B1) of protein G. This dual-color fluorescent QD probe binds the Fc region of immunoglobulin G (IgG) through its B1 domain at the QD surface and acts as a molecular-imaging probe at both the cellular and whole-body levels. In this paper, we present the synthesis of fluorescent, recombinant protein (HisEGFP-GB1)-conjugated, NIR-emitting QDs and their application to the dual-color molecular imaging of breast cancer cells in vitro and in vivo.
近红外(NIR)发射荧光探针广泛应用于全身水平的分子成像。然而,发射波长大于λ=700nm 的 NIR 发射荧光探针不适合用于细胞水平的分子成像,因为大多数常规荧光显微镜在近红外区域的光学灵敏度非常低。因此,为了实现使用单个探针进行细胞和全身水平的荧光成像,期望具有可见和 NIR 发射双色荧光探针。对于双色荧光分子成像,我们合成了荧光、重组蛋白偶联的 NIR 发射量子点(QD),其中重组蛋白由增强型绿色荧光蛋白(EGFP)和蛋白 G 的免疫球蛋白结合结构域(B1)组成。这种双色荧光 QD 探针通过其在 QD 表面的 B1 结构域与免疫球蛋白 G(IgG)的 Fc 区域结合,并作为细胞和全身水平的分子成像探针。在本文中,我们介绍了荧光、重组蛋白(HisEGFP-GB1)偶联的 NIR 发射 QD 的合成及其在体外和体内乳腺癌细胞的双色分子成像中的应用。