Campbell Charles T, Kim Gibum
Department of Chemistry, University of Washington, Seattle, Washington 98195-1700, USA.
Biomaterials. 2007 May;28(15):2380-92. doi: 10.1016/j.biomaterials.2007.01.047. Epub 2007 Mar 6.
Surface plasmon resonance (SPR) sensing has long been used to study biomolecular binding events and their kinetics in a label-free way. This approach has recently been extended to SPR microscopy, which is an ideal tool for probing large microarrays of biomolecules for their binding interactions with various partners and the kinetics of such binding. Commercial SPR microscopes now make it possible to simultaneously monitor binding kinetics on >1300 spots within a protein microarray with a detection limit of approximately 0.3 ng/cm(2), or <50 fg per spot (<1 million protein molecules) with a time resolution of 1s, and spot-to-spot reproducibility within a few percent. Such instruments should be capable of high-throughput kinetic studies of the binding of small ( approximately 200 Da) ligands onto large protein microarrays. The method is label free and uses orders of magnitude less of the precious biomolecules than standard SPR sensing. It also gives the absolute bound amount and binding stoichiometry.
表面等离子体共振（SPR）传感长期以来一直被用于以无标记的方式研究生物分子结合事件及其动力学。这种方法最近已扩展到SPR显微镜，它是探测生物分子大型微阵列与其各种配体的结合相互作用以及这种结合动力学的理想工具。现在，商业SPR显微镜能够同时监测蛋白质微阵列中1300多个位点上的结合动力学，检测限约为0.3 ng/cm²，或每个位点小于50 fg（小于100万个蛋白质分子），时间分辨率为1秒，位点间重现性在百分之几以内。此类仪器应能够对小分子（约200 Da）配体与大型蛋白质微阵列的结合进行高通量动力学研究。该方法无需标记，与标准SPR传感相比，使用的珍贵生物分子数量少几个数量级。它还能给出绝对结合量和结合化学计量。
