title: Accurate Quantitation of Dystrophin Protein in Human Skeletal Muscle Using Mass Spectrometry.
abstract: Quantitation of human dystrophin protein in muscle biopsies is a clinically relevant endpoint for both diagnosis and response to dystrophin-replacement therapies for dystrophinopathies. A robust and accurate assay would enable the use of dystrophin as a surrogate biomarker, particularly in exploratory Phase 2 trials. Currently available methods to quantitate dystrophin rely on immunoblot or immunohistochemistry methods that are not considered robust. Here we present a mass spectrometry based approach to accurately quantitate dystrophin protein in a total protein extract from human muscle biopsies. Our approach uses a combination of stable isotope labeled dystrophin as a spike-in standard, gel electrophoresis and high precision mass spectrometry to detect and quantitate multiple peptides of dystrophin within a complex protein mixture. The method was found highly reproducible and linear over a wide dynamic range, detecting as low as 5% of dystrophin relative to the normal amount in healthy individuals.
authors: Children's National Medical Center, Center for Genetic Medicine Research, USA ; Department of Integrative Systems Biology, The George Washington University, 2300 Eye Street, N.W., Ross 605, Washington, D.C. USA.
authors: Kristy J
authors: KJ
authors: Brown
authors: Ramya
authors: R
authors: Marathi
authors: Alyson A
authors: AA
authors: Fiorillo
authors: Eugene F
authors: EF
authors: Ciccimaro
authors: Seema
authors: S
authors: Sharma
authors: David S
authors: DS
authors: Rowlands
authors: Sree
authors: S
authors: Rayavarapu
authors: Kanneboyina
authors: K
authors: Nagaraju
authors: Eric P
authors: EP
authors: Hoffman
authors: Yetrib
authors: Y
authors: Hathout
keywords: Duchenne; Dystrophin; Mass spectrometry; Quantitation; Stable isotope
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