Steinkellner Hannes, Etzler Julia, Gmeiner Bernhard M, Laccone Franco
1 Institute of Medical Genetics, Medical University of Vienna , Vienna, Austria .
Assay Drug Dev Technol. 2015 Apr;13(3):167-73. doi: 10.1089/adt.2015.635. Epub 2015 Apr 7.
Spinal muscular atrophy (SMA) is a severe autosomal recessive disorder affecting one in every 10,000 live births. The disease is characterized by loss of alpha-motor neurons in the spinal cord that leads to progressive atrophy and weakness of limb and trunk muscles. This neuromuscular disorder results from deletions and/or mutations within the survival motor neuron 1 (SMN1) gene, leading to a pathologically decreased expression of functional full-length SMN protein. Here we report on the investigation to measure SMN protein levels through electrochemiluminescence immunoassay (ECLIA). This simple assay is a highly quantitative method able to measure SMN protein levels in human, mouse, and rat samples throughout a wide working range with low intra- and interassay error. The sensitivity for human SMN is 30 pg/mL and provides a new tool for the set up of high-throughput screening for basic research. Moreover, we describe a novel tool for a noninvasive assessment of SMN in buccal cells derived from healthy donors, SMA carriers, and SMA patients. The availability of a validated quantitative ECLIA should improve the investigation of novel compounds for the treatment of SMA.
脊髓性肌萎缩症（SMA）是一种严重的常染色体隐性疾病，每10000例活产中就有1例受影响。该疾病的特征是脊髓中的α运动神经元丧失，导致肢体和躯干肌肉进行性萎缩和无力。这种神经肌肉疾病是由生存运动神经元1（SMN1）基因的缺失和/或突变引起的，导致功能性全长SMN蛋白的病理性表达降低。在此，我们报告通过电化学发光免疫分析（ECLIA）测量SMN蛋白水平的研究。这种简单的分析方法是一种高度定量的方法，能够在很宽的工作范围内测量人、小鼠和大鼠样本中的SMN蛋白水平，且批内和批间误差低。人SMN的灵敏度为30 pg/mL，为基础研究的高通量筛选提供了一种新工具。此外，我们描述了一种用于非侵入性评估健康供体、SMA携带者和SMA患者颊细胞中SMN的新工具。经过验证的定量ECLIA的可用性应会改善对治疗SMA的新型化合物的研究。
