オーラルセッション
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2B-O1-0920 PDF
PAC-LC/MSを用いたマウス筋組織中ジストロフィンタンパク質の定量分析
The ability to quantify dystrophin protein as a biomarker is crucial in assessing the relationship between endogenous dystrophin protein expression and disease severity in Duchenne muscular dystrophy (DMD). We developed a highly sensitive peptide adsorption-controlled (PAC)-LC/MS-based method to quantify the absolute amount of endogenous dystrophin protein in mouse muscle lysates. To accurately quantify dystrophin protein, we determined quantitative values by the calibration curves using synthetic standard peptides and corrected them with tryptic digestion efficiency using partial length recombinant human dystrophin protein in mouse muscle lysate. Data showed that dystrophin expression in normal mouse and dystrophic mdx mouse muscle was 978 and 71.8 fmol/mg protein, respectively. Inter- and intra-assay precision were <15%. Our PAC-LC/MS method enables accurate quantification of dystrophin protein in mouse muscle with sufficient sensitivity, which can be useful for development of DMD therapy.