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Quantitative measurements of GDF‐8 using immunoaffinity LC‐MS/MS
Author(s) -
Palandra Joe,
Quazi Amira,
Fitz Lori,
Rong Haojing,
Morris Carl,
Neubert Hendrik
Publication year - 2016
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201500112
Subject(s) - gdf15 , antibody , chromatography , chemistry , monoclonal antibody , microbiology and biotechnology , biology , endocrinology , immunology
Purpose Growth and differentiation factor 8 (GDF‐8) is a negative regulator of skeletal muscle mass and targeted by inhibitors to treat diseases associated with muscle loss. In order to enable clinical and translational investigations of GDF‐8 inhibitors, specific and sensitive measurements of GDF‐8 are necessary. Experimental design An immunoaffinity LC‐MS/MS assay for quantification of GDF‐8 in serum was developed, qualified and implemented. The workflow includes offline enrichment of GDF‐8 using an anti‐GDF‐8 antibody, followed by isolation using magnetic beads, trypsin digestion, and quantification using 2D nanoflow LC‐MS/MS. Results This assay was qualified in human serum with a lower LOQ of 1.0 ng/mL based on the intact protein. GDF‐8 was quantified in serum from juvenile and adult humans as well as mouse, rat, and cynomolgus monkey. Additionally, the assay was utilized to demonstrate an increase of total GDF‐8 in serum following administration of an anti‐GDF‐8 monoclonal antibody therapeutic in cynomolgus monkeys. Conclusions and clinical relevance A specific and sensitive method was developed for the measurement of GDF‐8 in juvenile and adult serum samples as well as preclinical species. The confident quantification of GDF‐8 now enables a greater understanding of any association between changes GDF‐8 levels and muscle mass.