The development of biologics and biosimilars, which are produced by molecular and cell biology techniques, has been growing rapidly in recent decades. These are proteins among which recombinant monoclonal antibodies (mAbs) are gaining ground. Due to the long-term use and history of regulatory approvals, Chinese hamster ovary cell lines are most commonly used for the production of recombinant mAbs, in which transfection, amplification and clone selection is well established and characterized. Various analytical methods for monitoring critical quality attributes (CQA) are used during development of the cell line and bioprocess for the production of recombinant mAbs. The value of individual attribute must meet certain predefined criteria, as this is the only way to ensure adequate safety and efficacy of the therapeutic protein. Traditionally, specific analytical single-attribute methods are used for monitoring of CQA. Recently, however, traditional analytical methods have been replaced by the multi-attribute method (MAM), which is most often based on peptide mapping (Pepmap). Its main advantage over traditional analytical methods is monitoring of multiple attributes simultaneously in one analysis, thus saving a lot of time and resources. The main scope of our work was the establishment and optimization of MAM by using high-resolution liquid chromatography coupled with Orbitrap-based mass spectrometry technique. We have developed a useful tool for monitoring of multiple attributes simultaneously in one analysis, which could be an excellent alternative to conventional chromatographic and electrophoretic techniques.
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