Accurate Quantitation of Dystrophin Protein in Human Skeletal Muscle Using Mass Spectrometry
| Autor: | Sree Rayavarapu, Eugene F. Ciccimaro, Kristy J. Brown, Yetrib Hathout, David S. Rowlands, Seema Sharma, Kanneboyina Nagaraju, Alyson A. Fiorillo, Eric P. Hoffman, Ramya Marathi |
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| Rok vydání: | 2013 |
| Předmět: |
Gel electrophoresis
musculoskeletal diseases congenital hereditary and neonatal diseases and abnormalities biology Chemistry Pharmaceutical Science Skeletal muscle Exploratory phase Mass spectrometry Bioinformatics musculoskeletal system Article medicine.anatomical_structure Biochemistry biology.protein medicine Immunohistochemistry Biomarker (medicine) Dystrophin Total protein |
| Zdroj: | Journal of bioanalysisbiomedicine. |
| ISSN: | 1948-593X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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