Laboratory of Nanostructures and Biosensing, Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, Minnesota 55455, United States.
Anal Chem. 2012 Jul 17;84(14):6031-9. doi: 10.1021/ac300819a. Epub 2012 Jul 3.
Multiple sclerosis is a progressive neurological disorder that results in the degradation of myelin sheaths that insulate axons in the central nervous system. Therefore promotion of myelin repair is a major thrust of multiple sclerosis treatment research. Two mouse monoclonal natural autoantibodies, O1 and O4, promote myelin repair in several mouse models of multiple sclerosis. Natural autoantibodies are generally polyreactive and predominantly of the IgM isotype. The prevailing paradigm is that because they are polyreactive, these antibodies bind antigens with low affinities. Despite their wide use in neuroscience and glial cell research, however, the affinities and kinetic constants of O1 and O4 antibodies have not been measured to date. In this work, we developed a membrane biosensing platform based on surface plasmon resonance in gold nanohole arrays with a series of surface modification techniques to form myelin-mimicking lipid bilayer membranes to measure both the association and dissociation rate constants for O1 and O4 antibodies binding to their myelin lipid antigens. The ratio of rate constants shows that O1 and O4 bind to galactocerebroside and sulfated galactocerebroside, respectively, with unusually small apparent dissociation constants (K(D) ≈ 0.9 nM) for natural autoantibodies. This is approximately one to 2 orders of magnitude lower than typically observed for the highest affinity natural autoantibodies. We propose that the unusually high affinity of O1 and O4 to their targets in myelin contributes to the mechanism by which they signal oligodendrocytes and induce central nervous system repair.
多发性硬化症是一种进行性神经疾病,导致中枢神经系统轴突的髓鞘降解。因此,促进髓鞘修复是多发性硬化症治疗研究的主要方向。两种鼠源单克隆天然自身抗体 O1 和 O4 在几种多发性硬化症的小鼠模型中促进髓鞘修复。天然自身抗体通常具有多反应性,主要为 IgM 同种型。流行的观点是,由于它们具有多反应性,这些抗体与抗原的结合亲和力较低。然而,尽管它们在神经科学和神经胶质细胞研究中得到了广泛的应用,但迄今为止尚未测量 O1 和 O4 抗体的亲和力和动力学常数。在这项工作中,我们开发了一种基于金纳米孔阵列表面等离子体共振的膜生物传感器平台,通过一系列表面修饰技术形成髓鞘模拟脂质双层膜,以测量 O1 和 O4 抗体与髓鞘脂质抗原结合的结合和解离速率常数。速率常数的比值表明,O1 和 O4 分别与半乳糖脑苷脂和硫酸半乳糖脑苷脂结合,对于天然自身抗体来说,其表观解离常数(K(D)≈0.9 nM)非常小。这大约比通常观察到的最高亲和力天然自身抗体低 1 到 2 个数量级。我们提出,O1 和 O4 对髓鞘中靶标的异常高亲和力有助于它们向少突胶质细胞发出信号并诱导中枢神经系统修复的机制。