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Ara h 1 protein-antibody dissociation study: evidence for binding inhomogeneities on a molecular scale

机译：Ara h 1蛋白抗体解离研究：在分子水平上结合不均一性的证据

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The characterization of biomolecular interactions is essential when designing novel biosensors, since the interaction between the bioreceptor and the ligand determines important biosensing parameters such as sensitivity and selectivity. In this paper we study the interaction of the trimeric Ara h 1 protein with a monoclonal anti-Ara h 1 antibody by means of magnetic force-induced dissociation. The proteins were bound to magnetic particles and polystyrene surfaces by EDC/NHS reaction chemistry and by physisorption, respectively. Two different molecular configurations have been investigated, with either the Ara h 1 protein on the particles or the Ara h 1 protein on the polystyrene surface. A model with a Gaussian distribution of energy barriers for dissociation gives an adequate description for the measured multi-exponential decays. We hypothesize that distributions of molecular orientations as well as experimentally induced variations may underlay the observed distributions. The two molecular configurations show a different peak value of the energy distribution. Similarly, SPR experiments for two distinct configurations (either Ara h 1 protein on the surface, or anti-Ara h 1 antibody on the surface) also show clear differences in dissociation behavior. We hypothesize that the multivalency of the involved molecules leads to different modes of binding. The results of this work highlight the importance of molecular inhomogeneities when studying the interaction processes of biomolecular complexes. Research Paper
机译：设计新型生物传感器时，生物分子相互作用的表征至关重要，因为生物受体与配体之间的相互作用决定了重要的生物传感参数，例如灵敏度和选择性。在本文中，我们通过磁力诱导的解离研究了三聚体Ara h 1蛋白与单克隆抗Ara h 1抗体的相互作用。分别通过EDC / NHS反应化学和物理吸附将蛋白质结合到磁性颗粒和聚苯乙烯表面。已经研究了两种不同的分子构型，即颗粒上的Ara h 1蛋白或聚苯乙烯表面上的Ara h 1蛋白。具有用于解离的能垒的高斯分布的模型为所测量的多指数衰减提供了足够的描述。我们假设分子取向的分布以及实​​验诱导的变化可能是观察到的分布的基础。两种分子构型显示出不同的能量分布峰值。同样，针对两种不同构型（表面上的Ara h 1蛋白或表面上的抗Ara h 1抗体）的SPR实验也显示出解离行为的明显差异。我们假设所涉及分子的多价导致不同的结合模式。这项工作的结果突出了研究生物分子复合物相互作用过程时分子不均一性的重要性。研究论文

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