Simultaneous detection method of muscle developing monoclonal antibodies with using LC-MS/MS from Dried Blood Spots and Plasma

Abstract

Current treatments aim to slow muscle degeneration through diet, while manipulating TGF-β family pathways, such as myostatin, activin A, and GDF-11, offers potential for addressing muscle wasting. However, the use of inhibitors to enhance athletic performance is prohibited by WADA. This study developed a method to detect monoclonal antibodies such as Landogrozumab, Domagrozumab, and Bimagrumab, specifically targeting the TGF-β superfamily. To confirm assay stability, we included Dulaglutide as an internal standard. Utilizing Both Dried blood spots and Plasma, antibodies were purified using protein G magnetic beads to minimize interference, followed by LC-MS/MS analysis. The validation was conducted specificity, selectivity, linearity, precision, recovery, matrix effect. All targeted monoclonal antibodies were below the limit of detection <0.5 ？g/ml. The assays showed strong linearity (R2=0.99). No interfering signals were detected that could affect the detection of the targets and Carryover was not detected. Since the matrix effect for most monoclonal antibodies is nearly 100%, using both samples in this multiplex analysis approach is appropriate. Though anti-TGF-β superfamily antibodies lack clinical approval, detecting prohibited compounds is essential for doping control. Also, using DBS for doing analysis offers a fast, safe, and stable process for athletes and analysts. We aimed to develop and validate a multiplex method for detecting prohibited monoclonal antibodies in plasma and DBS.