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用单细胞离心法测试生物分子相互作用的动力强度

Hans T Bergal^1,2, Koji Kinoshita^1,2, Wesley P Wong^1,2,3,4

¹Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, Massachusetts 02215, United States.

ACS central science

|October 27, 2025

概括

一种新的离心力显微镜 (CFM) 能够高通量测量分子结合力. 这种方法量化了受体-连接体相互作用和T细胞参与治疗开发.

科学领域:

生物物理化学生物物理化学
分子识别分子识别
免疫学免疫学免疫学

背景情况:

受体-连接体相互作用对于免疫监测和组织发育等生物过程至关重要.
现有的研究结合热情的方法缺乏高吞吐量或精度.
了解分子结合动力学对于治疗开发至关重要.

研究的目的:

开发一种高通量方法来量化分子和细胞结合动力学.
用双通道光成像来增强现有的离心力显微镜 (CFM).
为了描述双特异性T细胞参与剂 (BiTE) 分子的结合机制.

主要方法:

利用离心力显微镜 (CFM) 对数千个单细胞进行实时并行力测量.
开发了一种下一代CFM,具有双通道光成像,以跟踪单个细胞解结事件.
进行了细胞蛋白和细胞细胞测试,以量化欲和表征BiTE介导的粘附.

主要成果:

量化了与BiTE修饰的表面相互作用的T细胞和B细胞的狂热度,显示了连接体度和键强度之间的受体特异性相关性.
在Jurkat和Nalm6细胞之间表现出BiTE介导的粘附性,显示出依赖时间的狂热度增加.
证明了CFM研究受体介导相互作用的机械化学原理的能力.

结论:

集成的CFM提供了一种高通量方法,用于研究受体-连接体相互作用和结合性.
这种方法对生物物理化学,分子识别和免疫疗法的发展有着广泛的影响.
CFM有助于更深入地了解细胞粘附和免疫反应的机械化学基础.

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