High-throughput untargeted screening of biotherapeutic macromolecules in equine plasma by UHPLC-HRMS/MS: Application to monoclonal antibodies and Fc-fusion proteins for doping control.

This study focuses on high-throughput untargeted screening of biotherapeutic macromolecules in equine plasma by uhplc-hrms/ms: application to monoclonal antibodies and fc-fusion proteins for doping control.. The research employs high-performance liquid chromatography (HPLC) techniques to address analytical challenges in the biopharmaceutical field. Many innovative biotherapeutics have been marketed in the last decade. Monoclonal antibodies (mAbs) and Fc-fusion proteins (Fc-proteins) have been developed for the treatment of diverse diseases (cancer, autoimmune diseases, and inflammatory disorders) and now represent an important part of targeted therapies. However, the ready availability of such biomolecules, sometimes characterized... Research Background and Significance The rapid expansion of biotherapeutics, especially monoclonal antibodies (mAbs) and Fc-fusion proteins, has revolutionized targeted therapies for a range of diseases including cancer, autoimmune disorders, and inflammatory conditions. These large and structurally complex macromolecules present unique analytical challenges due to their size, heterogeneity, and post-translational modifications. In equine medicine and doping control, the detection and monitoring of such biotherapeutics become critical to ensure fair competition and animal welfare. The study by Pinetre et al. addresses an urgent need for robust, high-throughput, untargeted screening methodologies capable of sensitively detecting these molecules directly in equine plasma. Utilizing ultrahigh-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UHPLC-HRMS/MS), this work provides a powerful platform that overcomes limitations of specificity, throughput, and sensitivity encountered by conventional assays. The