Chair of Experimental Physics & Center for NanoScience, Ludwig-Maximilians-University München, Munich, Germany.
Chair of Experimental Physics & Center for NanoScience, Ludwig-Maximilians-University München, Munich, Germany.
Biophys J. 2013 Dec 17;105(12):2687-94. doi: 10.1016/j.bpj.2013.10.013.
Gaining information about receptor profiles on cells, and subsequently finding the most efficient ligands for these signaling receptors, remain challenging tasks in stem cell and cancer research as well as drug development. We introduce a live-cell method with great potential in both screening for surface receptors and analysing binding forces of different ligands. The technique is based on the molecular force assay, a parallel-format, high-throughput experiment on a single-molecule level. On human red blood cells, we demonstrate the detection of the interaction of N-acetyl-α-D-galactosaminyl residues with the lectin helix pomatia agglutinine and of the CD47 receptor with its antibody. The measurements are performed under nearly physiological conditions and still provide a highly specific binding signal. Moreover, with a detailed comparative force analysis on two cell types with different morphology, we show that our method even allows the determination of a DNA force equivalent for the interaction of the CD47 receptor and its antibody.
获取细胞表面受体的信息,并找到这些信号受体的最有效配体,这仍然是干细胞和癌症研究以及药物开发中的挑战性任务。我们引入了一种活细胞方法,在筛选表面受体和分析不同配体的结合力方面具有很大的潜力。该技术基于分子力测定法,这是一种在单细胞水平上进行的平行格式、高通量实验。我们在人红细胞上演示了 N-乙酰-α-D-半乳糖胺残基与凝集素舞毒蛾凝集素相互作用以及 CD47 受体与其抗体相互作用的检测。测量是在近乎生理的条件下进行的,但仍提供了高度特异性的结合信号。此外,通过对两种形态不同的细胞类型进行详细的比较力分析,我们表明,我们的方法甚至可以确定 CD47 受体与其抗体相互作用的 DNA 力等效值。