Analysis of Nonhuman N-Glycans as the Minor Constituents in Recombinant Monoclonal Antibody Pharmaceuticals
| Autor: | Soichiro Kita, Takao Hayakawa, Eiki Maeda, Koji Urakami, Kazuaki Kakehi, Mitsuhiro Kinoshita |
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| Rok vydání: | 2012 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Glycan Glycosylation medicine.drug_class Electrospray ionization Molecular Sequence Data Monoclonal antibody Epitope Analytical Chemistry law.invention chemistry.chemical_compound Polysaccharides law medicine Chromatography High Pressure Liquid Chromatography biology Lasers Immunogenicity Antibodies Monoclonal Electrophoresis Capillary Galactose Recombinant Proteins Carbohydrate Sequence chemistry Biochemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Recombinant DNA Neuraminic Acids Antibody |
| Zdroj: | Analytical Chemistry. 84:2373-2379 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac300234a |
| Popis: | Minor N-linked glycans containing N-glycolylneuraminic acid residues and/or α-Gal epitopes (i.e., galactose-α1,3-galactose residues) have been reported to be present in recombinant monoclonal antibody (mAb) therapeutics. These contaminations are due to their production processes using nonhuman mammalian cell lines in culture media containing animal-derived materials. In case of the treatment of tumors, we inevitably use such mAbs by careful risk-benefit considerations to prolong patients' lives. However, expanding their clinical applications such as for rheumatism, asthma, and analgesia demands more careful evaluation of the product characteristics. The present work for detailed evaluations of N-glycans demonstrates the methods using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) and a combination of high-performance liquid chromatography and electrospray ionization time-of-flight mass spectrometry. The CE-LIF method provides excellent separation of both major and minor N-glycans from six commercial mAb pharmaceuticals within 30 min and clearly indicates that a possible trigger of immunogenicity in humans due to the presence of nonhuman N-glycans is present. We strongly believe that the proposed method will be a powerful tool for the analysis of N-glycans of recombinant mAb products in various development stages, such as clone selection, process control, and routine release testing to ensure safety and efficacy of the products. |
| Databáze: | OpenAIRE |
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