Elucidation of Degradants in Acidic Peak of Cation Exchange Chromatography in an IgG1 Monoclonal Antibody Formed on Long-Term Storage in a Liquid Formulation
| Autor: | Pavel V. Bondarenko, Sejal Gandhi, Sampathkumar Krishnan, Gang Xiao, Da Ren, Christopher James Sloey, Margaret Speed Ricci |
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| Rok vydání: | 2011 |
| Předmět: |
Glycosylation
medicine.drug_class Chemistry Pharmaceutical Drug Storage Glutamine Ion chromatography Size-exclusion chromatography Pharmaceutical Science CHO Cells Receptors Fc Monoclonal antibody Peptide Mapping Mass Spectrometry chemistry.chemical_compound Drug Stability Glycation Cricetinae medicine Animals Humans Trypsin Pharmacology (medical) Deamidation Pharmacology Aspartic Acid Chromatography Reverse-Phase Chromatography Isoelectric focusing Chemistry Histocompatibility Antigens Class I Organic Chemistry Temperature Antibodies Monoclonal Biological activity Chromatography Ion Exchange Complementarity Determining Regions Biochemistry Immunoglobulin G Chromatography Gel Molecular Medicine Asparagine Isoelectric Focusing Chromatography Liquid Biotechnology |
| Zdroj: | Pharmaceutical Research. 29:209-224 |
| ISSN: | 1573-904X 0724-8741 |
| Popis: | An IgG1 therapeutic monoclonal antibody showed an increase in acidic or pre-peak by cation exchange chromatography (CEX) at elevated temperatures, though stable at 2–8°C long-term storage in a liquid formulation. Characterization effort was undertaken to elucidate the degradants in CEX pre-peak and effect on biological activity. Purified CEX fractions were collected and analyzed by peptide mapping, size exclusion, intact and reduced-alkylated reversed phase techniques. Biophysical characterization, isoelectric focusing and Isoquant analysis were also performed to determine nature of degradants. Bioassay and surface plasmon resonance experiments were performed to determine the impact on biological activity of the degradants. No major degradation due to oxidation, clipping or aggregation was detected; conformational differences between purified fractions observed were not significant. Sialic acid, N-terminal glutamine cyclization and glycation differences contributed to the CEX pre-peak in the mAb control sample; increase in CEX pre-peak at 25°C and higher was caused by additive degradation pathways of deamidation, related isomerization and clipping. The observed CEX pre-peak increase was caused by multiple degradations, especially deamidation and clipping. This elucidation of degradants in CEX peaks may apply to other therapeutic IgG1 monoclonal antibodies. |
| Databáze: | OpenAIRE |
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