Kim Lina, Jo Seongjae, Kim Gyeong-Ji, Kim Kyung Ho, Seo Sung Eun, Ryu Eunsu, Shin Chan Jae, Kim Yu Kyung, Choi Jeong-Woo, Kwon Oh Seok
Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Republic of Korea.
Department of Chemical & Biomolecular Engineering, Sogang University, Seoul, 04107, Republic of Korea.
Nano Converg. 2023 Oct 30;10(1):51. doi: 10.1186/s40580-023-00399-x.
Antibody sensor to detect viruses has been widely used but has problems such as the difficulty of right direction control of the receptor site on solid substrate, and long time and high cost for design and production of antibodies to new emerging viruses. The virus detection sensor with a recombinant protein embedded liposome (R/Li) was newly developed to solve the above problems, in which R/Li was assembled on AuNPs (Au@R/Li) to increase the sensitivity using localized surface plasmon resonance (LSPR) method. Recombinant angiotensin-converting enzyme-2 (ACE2) was used as host receptors of SARS-CoV and SARS-CoV-2, and the direction of enzyme active site for virus attachment could be controlled by the integration with liposome. The recombinant protein embedded liposomes were assembled on AuNPs, and LSPR method was used for detection. With the sensor platform S1 protein of both viruses was detected with detection limit of 10 pg/ml and SARS-CoV-2 in clinical samples was detected with 10 ~ 35 Ct values. In the selectivity test, MERS-CoV did not show a signal due to no binding with Au@R/Li. The proposed sensor platform can be used as promising detection method with high sensitivity and selectivity for the early and simple diagnosis of new emerging viruses.
用于检测病毒的抗体传感器已被广泛使用,但存在诸如难以控制固体基质上受体位点的正确方向,以及针对新出现病毒设计和生产抗体所需时间长且成本高等问题。为了解决上述问题,新开发了一种带有重组蛋白嵌入脂质体(R/Li)的病毒检测传感器,其中R/Li组装在金纳米颗粒(Au@R/Li)上,以使用局域表面等离子体共振(LSPR)方法提高灵敏度。重组血管紧张素转换酶2(ACE2)被用作严重急性呼吸综合征冠状病毒(SARS-CoV)和严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的宿主受体,并且病毒附着的酶活性位点的方向可以通过与脂质体整合来控制。将重组蛋白嵌入脂质体组装在金纳米颗粒上,并使用LSPR方法进行检测。利用该传感器平台检测到两种病毒的S1蛋白,检测限为10 pg/ml,并且检测到临床样本中的SARS-CoV-2,Ct值为10至35。在选择性测试中,中东呼吸综合征冠状病毒(MERS-CoV)由于未与Au@R/Li结合而未显示信号。所提出的传感器平台可作为一种有前景的检测方法,用于对新出现病毒进行高灵敏度和高选择性的早期简易诊断。