Articles tagged ”mAb Quality”
mAb Deamidation Study using FabRICATOR® Digestion and HIC Separation
Hydrophobic interaction chromatography (HIC) is often used in characterization of therapeutic antibody products due to its ability to separate direct or indirect structural changes in the studied protein. Scientists at Alexion have published a study where FabRICATOR (IdeS) was used to generate Fc and F(ab’)2 fragments of an antibody to study conformational changes of a monoclonal antibody (King et al. 2018).
Separation of the intact antibody on HIC reveled two major peaks that were collected and subjected to FabRICATOR digestion. After digestion, the Fc and F(ab’)2 fragments separated well, and the heterogeneity was localized to the F(ab’)2 domain. Variations in the Fc were observed and attributed to oxidation modifications. Peptide mapping of the domains were carried out and a 1 Da difference was localized , indicating deamidation of Asn to either Asp or isoAsp in the complementarity-determining region (CDR) of the light chain. The observed difference in HIC separation pattern was also linked to changes in antigen binding, since the deamidation of the Asn residues reduced the binding of the antibody to its target antigen.
Taken together, this paper indicates that a single deamidation in the light chain changed the hydrophobicity profile of the antibody and impacted the antigen binding. The use of FabRICATOR (IdeS) digestion and HIC separation could serve as a quick screening assay to study deamidation changes in the F(ab’)2 domain.
Real-time Monitoring of Antibody Quality Control using FabRICATOR
The production of antibody-based therapeutics requires careful monitoring of quality attributes. Researchers at Amgen developed a technology to model and monitor the high-mannose content of the antibody in real-time using FabRICATOR digestion (Zupke et al. 2015). Read more »
Study of Antibody Quality Attributes on Cell Culture Supernatant
The production of antibody-based therapeutics requires careful monitoring of quality attributes. Researchers at Institute Pasteur in collaboration with LFB Biotechnologies have used FabRICATOR to monitor important quality attributes of a therapeutic antibody, directly on the cell culture supernatant (Henninot et al. 2015).
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