Pavlickova Petra, Jensen Nina Mejlhede, Paul Hubert, Schaeferling Michael, Giammasi Chiara, Kruschina Margit, Du Wei-Dong, Theisen Michael, Ibba Michael, Ortigao Flavio, Kambhampati Dev
ThermoHybaid, Sedanstrasse 18, 890 77 Ulm, Germany.
J Proteome Res. 2002 May-Jun;1(3):227-31. doi: 10.1021/pr0200036.
We report a novel high-throughput (HTP) protein chip platform, constructed on gold using self-assembly techniques, for conducting high quality antigen-antibody interactions. Biotinylated monolayers were used to immobilize a streptavidin surface with high packing density. This biocompatible platform was then used for detection of serum IgM antibodies. Serum samples of patients suspected to suffer from Lyme borreliosis were used to validate the protein chip platform using biotinylated peptide AAOspC8 molecules as the test probes. Various experimental parameters such as the effect of concentration of probes, targets, temperature of incubation, and their effect on the resulting signal-to-noise ratio are described in detail. Highly specific protein interaction data with a high signal-to-noise ratio were obtained with serum sample solutions as low as 1 microL/spot (1/10 diluted).
我们报道了一种新型的高通量（HTP）蛋白质芯片平台，该平台利用自组装技术构建于金表面，用于进行高质量的抗原-抗体相互作用。生物素化单层用于以高堆积密度固定链霉亲和素表面。然后将这个生物相容性平台用于检测血清IgM抗体。疑似患有莱姆病的患者的血清样本被用于以生物素化肽AAOspC8分子作为测试探针来验证蛋白质芯片平台。详细描述了各种实验参数，如探针浓度、靶标浓度、孵育温度的影响及其对所得信噪比的影响。使用低至1微升/点（1/10稀释）的血清样本溶液获得了具有高信噪比的高度特异性蛋白质相互作用数据。
