Université Paris-Saclay, Univ Evry, CNRS, LAMBE, Evry-Courcouronnes, Paris, France.
CY Cergy Paris Université, CNRS, LAMBE, Cergy, Paris, France.
Methods Mol Biol. 2021;2237:55-67. doi: 10.1007/978-1-0716-1064-0_5.
The coupling of surface plasmon resonance imaging (SPRi) with mass spectrometry (MS) offers a very promising multidimensional analysis. This system takes advantage of the two well-established techniques: SPR, which allows for the analysis of biomolecular interactions through the determination of kinetic and thermodynamic constants, and MS, which can characterize biological structures from mass measurements and fragmentation experiments. Here, a protocol for the coupling of SPRi with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is described using a biochip grafted by antibodies in an array format. Interaction between β-lactoglobulin antibodies and the protein antigen is detected and analyzed by SPRi. Then, the arrayed biochip which fitted a commercially MALDI target was inserted in a MALDI source, and mass spectra were recorded directly from the biochip surface from each antibody spot, showing protein ions attributed to the corresponding specific protein antigens.
表面等离子体共振成像（SPRi）与质谱（MS）的结合提供了一种非常有前途的多维分析。该系统利用了两种成熟的技术：SPR，它允许通过确定动力学和热力学常数来分析生物分子相互作用，以及 MS，它可以通过质量测量和碎片实验来表征生物结构。本文描述了一种使用阵列形式接枝抗体的生物芯片将 SPRi 与基质辅助激光解吸/电离质谱（MALDI-MS）相结合的方案。通过 SPRi 检测和分析β-乳球蛋白抗体与蛋白质抗原之间的相互作用。然后，将适合商用 MALDI 靶的阵列生物芯片插入 MALDI 源中，直接从每个抗体点的生物芯片表面记录质谱，显示归因于相应特定蛋白质抗原的蛋白质离子。
