Characterization of Pharmaceutical IgG and Biosimilars Using Miniaturized Platforms and LC-MS/MS

Abstract

Therapeutic monoclonal antibodies (mAbs) have made a tremendous impact in treating patients with various diseases. MAbs are designed to target specifically a cell and illicit a response from the immune system to destroy the cell. As originator mAb drug patents are coming to an end, generic pharmaceutical companies are poised to replicate and produce so-called biosimilar drugs. MAbs are significantly more complicated than small drugs to analyze and produce. The mAb proteoform and glycoform must be as similar to the original drug as possible to be a viable replacement. The mAb proteoform is well characterized but can be altered through various undesirable reactions such as deamidation. The mAb glycoform is harder to replicate as the glycan formation is a complicated template-less one; it is proving difficult for the originator companies to produce a homogenous population of mAbs from batch to batch. Severe side-effects have occurred in patients taking mAbs with immunogenic glycans, highlighting the importance of quality control mechanisms. The complex nature of mAbs requires sensitive and robust tools amenable to the high-throughput analysis required by a manufacturing setting. Miniaturized analytical platforms for complex biosimilar analysis are still in their infancy but have shown great promise for sample preparation. Capillary electrophoresis-laser induced fluorescence remains a powerful and fast technique for routine glycan analysis. Mass spectrometry is the method of choice for the analysis of mAb proteoforms and is emerging as a powerful tool for glycoform analysis.

A systematic approach for peptide characterization of B-cell receptor in chronic lymphocytic leukemia cells

“…Related to this, the production of biosimilars (biologic medical products which are almost identical copies of original products), which are considered as good alternatives to improve healthcare access and outcomes at reduced costs, could be greatly improved if disease-related antibody sequences are determined (e.g., monoclonal antibodies, cytokines…) [22, 23]. …”

Characterization of isomeric glycan structures by LC‐MS/MS

“…Thus, the basic conditions of HPAEC allow efficient separation of glycans based on their formal charges, sizes, monosaccharide compositions, and intramolecular linkages . Oligosaccharides and glycoforms of glycoproteins have been demonstrated to be essential for many biological functions, medicines, and diseases . Therefore, there is a continued need for better approaches to analyze glycans.…”

LC–MS/MS glycomics of idiopathic rapid eye movement sleep behavior disorder

“…Recently, adopting a proteomics‐driven discovery analysis combined with network‐based approach we have identified novel potential actionable protein biomarkers embedded in RBD pathogenesis and molecular pathways , However, there is now substantial evidence that glycosylation, an important post‐translational modification, influences the standard stability and functions of many proteins and alter their downstream biological processes. As such, it has also been implicated in dynamic disease mechanisms and the onset of many pathological conditions , but no data are available for RBD. We hypothesized that the characterization of the glycosylation pattern of patients with RDB would be of great value to understand the pathophysiology and underlying mechanisms and represent potentially useful biomarkers for disease‐associated molecular changes.…”