建立等离子共振/质谱矩阵芯片平台

Protein microarrays are the format of choice for high throughput, high content protein interaction analysis. In most of the array formats reporter molecules are used in multistep detection of the protein interactions. Among the few exixiting lable-free detection approaches, surface plasmon resonance(SPR) and mass spectormetry (MS) stand out as most promising for utilization in protein microarrays, albeit both have been used only sporadically for high content protein arrays. Shown here for the first time is the combination of SPR and MS detections on a single high content protein microarry. Antibodies to five human plasma proteins were arrayed in a 10 x 10 spot arrangement on a chemically activated gold coated glass chip. Binding of proteins to their corresponding antibodies was monitored via SPR imaging across the entire surface of the chip. Following protein affinity retrieval, the chip was overlaid with MALDI matrix and MS analyzed, producing protein-specific mass spectra from distinct spots on the array. The SPR/MS dual detection is well suited for high content protein microarrays and comprehensive protein analysis-from quantitative assessment of the protein concentration to detection of structural protein variants arising from genetic variations and postexpression procedding.