Gao Yingying, Liu Yueling, Yan Rui, Zhou Jinfeng, Dong Hao, Hua Xin, Wang Ping
State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai 200237, P. R. China.
Key Laboratory of Advanced Materials, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, P. R. China.
Anal Chem. 2020 Oct 6;92(19):13595-13603. doi: 10.1021/acs.analchem.0c03476. Epub 2020 Sep 17.
Real-time in situ imaging of organelles is increasingly important in modern biomedical analysis and diseases diagnosis. To realize this goal, organelle-targeting nanoparticles as one of the most commonly used technologies in subcellular sensing and imaging has attracted a lot of interest. The biocompatibility, specificity, and binding efficiency are especially critical for efficient organelle-targeting bioimaging. Gold nanoparticles (AuNPs) fabricated with bifunctional peptides constructed with both Au-binding affinity and nucleus-targeting ability were designed and examined for efficient nucleus-targeting bioimaging. Such a design is expected to achieve an oriented assembling of peptides by the medium of the Au-binding peptides specifically assembled on the surface of AuNPs, with the nucleus-targeting end open for accessibility. The bifunctional peptides showed strong binding affinity toward AuNPs and led to a binding capability ∼1.5 times higher than that of the bare nucleus-targeting peptides, ensuring good surface coverage of the nanoparticles for enhanced nucleus-targeting ability. Such fabricated AuNPs demonstrated over 90% cell viability after incubation for 24 h with HepG2 cells, which were highly biocompatible. Precise and efficient bioimaging of the nucleus was achieved for HepG2 cells by using the fabricated AuNPs as observed with a confocal laser scanning microscope, a dark-field/fluorescence microscope, and a transmission electron microscope. The high surface coverage and oriented binding pattern appeared to be a promising strategy for construction of organelle-targeting agencies.
细胞器的实时原位成像在现代生物医学分析和疾病诊断中变得越来越重要。为实现这一目标，细胞器靶向纳米颗粒作为亚细胞传感和成像中最常用的技术之一，已引起广泛关注。生物相容性、特异性和结合效率对于高效的细胞器靶向生物成像尤为关键。设计并研究了用具有金结合亲和力和细胞核靶向能力的双功能肽制备的金纳米颗粒（AuNPs）用于高效的细胞核靶向生物成像。这种设计有望通过特异性组装在AuNPs表面的金结合肽为媒介实现肽的定向组装，使细胞核靶向端开放以易于接近。双功能肽对AuNPs表现出很强的结合亲和力，其结合能力比单纯的细胞核靶向肽高出约1.5倍，确保了纳米颗粒良好的表面覆盖率，从而增强了细胞核靶向能力。用这种制备的AuNPs与HepG2细胞孵育24小时后，细胞活力超过90%，表明其具有高度的生物相容性。使用制备的AuNPs，通过共聚焦激光扫描显微镜、暗场/荧光显微镜和透射电子显微镜观察，实现了对HepG2细胞核的精确高效生物成像。高表面覆盖率和定向结合模式似乎是构建细胞器靶向制剂的一种有前景的策略。
