Identification of leucocyte surface protein interactions by high-throughput screening with multivalent reagents
Identification of leucocyte surface protein interactions by high-throughput screening with multivalent reagents
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DOI：
10.1111/j.1365-2567.2009.03153.x
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发表时间：
2010-01-01
期刊：
影响因子：
6.4
通讯作者：
Barclay, A. Neil
中科院分区：
文献类型：
--
作者：
Jiang, Lei;Barclay, A. Neil
我们描述了一种高通量筛选系统来检测白细胞表面蛋白之间的相互作用，考虑到这些相互作用通常是非常低的亲和力。这种方法包括生产带有标签的白细胞蛋白质的胞外区域，以便它们可以与纳米颗粒结合，提供亲和试剂，以筛选使用Proteon(TM)XPR36固定化的36种类似蛋白质，并通过表面等离子共振进行检测。该系统使用已建立的相互作用进行测试，这些相互作用可以在没有虚假绑定的情况下被检测到。通过鉴定癌胚抗原相关细胞黏附分子1和癌胚抗原相关细胞黏附分子8之间的新相互作用，显示了检测新相互作用的能力。
P>We describe a high-throughput screening system to detect interactions between leucocyte surface proteins, taking into account that these interactions are usually of very low affinity. The method involves producing the extracellular regions of leucocyte proteins with tags so that they can be bound to nanoparticles to provide an avid reagent to screen over an array of 36 similar proteins immobilized using the Proteon (TM) XPR36 with detection by surface plasmon resonance. The system was tested using established interactions that could be detected without spurious binding. The ability to detect new interactions was shown by identifying a new interaction between carcinoembryonic antigen-related cell adhesion molecule 1 and carcinoembryonic antigen-related cell adhesion molecule 8.