AGADA BioSciences Inc., Halifax, Nova Scotia B3H0A8, Canada.
Altasciences, Laval, Québec H7V4B3, Canada.
Biomark Med. 2019 Oct;13(14):1209-1225. doi: 10.2217/bmm-2019-0242. Epub 2019 Aug 5.
Detection of drug-induced dystrophin in patient muscle biopsy is a surrogate outcome measure for Duchenne muscular dystrophy. We sought to establish and validate an orthogonal approach to measurement of dystrophin protein and RNA in muscle biopsies. Validated methods were developed for dystrophin western blotting, mass spectrometry, immunostaining and reverse transcriptase PCR of biopsy mRNA using muscle biopsy standards. Both western blotting and mass spectrometry validated methods demonstrated good linearity, and acceptable precision and accuracy with a lower limit of quantitation at 1%. Immunostaining and reverse transcriptase PCR methods were shown to be reliable. The described orthogonal approach is sufficient to support measures of dystrophin as a surrogate outcome in a clinical trial.
在患者肌肉活检中检测药物诱导的抗肌萎缩蛋白是杜氏肌营养不良症的替代终点指标。我们试图建立和验证一种用于测量肌肉活检中抗肌萎缩蛋白蛋白和 RNA 的正交方法。针对肌肉活检标准，开发了用于抗肌萎缩蛋白 Western 印迹、质谱、免疫染色和逆转录聚合酶链反应的验证方法。Western 印迹和质谱验证方法均显示出良好的线性，可接受的精密度和准确度，定量下限为 1%。免疫染色和逆转录聚合酶链反应方法被证明是可靠的。所描述的正交方法足以支持将抗肌萎缩蛋白作为临床试验替代终点的测量。
