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Comparing the original and biosimilar biotherapeutics of the monoclonal antibody eculizumab by intact mass measurement and middle-up mass spectrometry analysis

https://doi.org/10.32362/2410-6593-2021-16-1-76-87

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Abstract

Objectives. In this biosimilar research, we compare the monoclonal antibody eculizumab obtained from different drugs [original Soliris® (Alexion Pharmaceuticals) and candidate Elizaria® (Generium)] by intact mass measurement and middle-up mass spectrometry analysis to enhance the role of mass spectrometry methods in biopharmaceutical development processes.

Methods. The intact mass measurement is performed using a high-resolution ESI-MS. The middle-up analysis is performed by reversed-phase high-performance liquid chromatography with ESI-MS detection, subsequent IdeS treatment of antibodies, and disulfide bond reduction.

Results. We have shown some small differences between the original and candidate drugs in the minor glycans level. Man5 glycan is only found in the original Soliris, and G0 is only found in the Elizaria. Glycation sites are also found in the light chain and Fd subunits of the original Soliris. The glycation level does not exceed 4.4%. The non-clipped C-end lysine level and G0F glycan levels are slightly lower in the original Soliris. All registered differences are not crucial for eculizumab’s quality and do not affect its effectiveness and preclinical safety. Generally, the results show a high level of similarity between the original and candidate drugs.

Conclusions. The comparative mass spectrometry analysis of eculizumab in the original Soliris and Elizaria allows us to estimate their high degree of similarity by molecular masses and major modification profiles.

About the Authors

M. B. Degterev
International Biotechnology Center Generium
Russian Federation

Maksim B. Degterev, Research Scientist, Physico-Chemical Methods Laboratory

14, Vladimirskaya ul., Volginskiy, Vladimir oblast, 601125



R. R. Shukurov
International Biotechnology Center Generium
Russian Federation

Rakhim R. Shukurov, Head of the Analytical Methods Department

14, Vladimirskaya ul., Volginskiy, Vladimir oblast, 601125



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Supplementary files

1. Fig. 1. Mechanism of action of the IdeS protease.
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2. This is to certify that the paper Comparing the original and biosimilar biotherapeutics of the monoclonal antibody eculizumab by intact mass measurement and middle-up mass spectrometry analysis commissioned to us by Maksim B. Degterev, Rakhim R. Shukurov has been edited for English language and spelling by Enago, an editing brand of Crimson Interactive Inc.
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  • We have conducted a mass spectrometric study of the comparability of the original and biosimilar preparations of the monoclonal antibody eculizumab at the levels of intact molecules and subunits after IdeS proteolysis and restoration of disulfide bonds.
  • At the level of intact molecules, several differences in glycosylation profiles are found. However, the study of subunits shows that a significant part of them is due to the presence of additional glycation sites in the LC and Fd subunits of eculizumab of the original drug.
  • Mass spectrometric analysis of samples of the original and biosimilar preparations of the monoclonal antibody eculizumab at the intact molecules and subunits levels shows a high comparability level.

For citation:


Degterev M.B., Shukurov R.R. Comparing the original and biosimilar biotherapeutics of the monoclonal antibody eculizumab by intact mass measurement and middle-up mass spectrometry analysis. Fine Chemical Technologies. 2021;16(1):76-87. https://doi.org/10.32362/2410-6593-2021-16-1-76-87

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ISSN 2410-6593 (Print)
ISSN 2686-7575 (Online)