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Updated: Jan 23, 2026

A Fast and Quantitative Method for Post-translational Modification and Variant Enabled Mapping of Peptides to Genomes
Published on: May 22, 2018
Native peptide mapping - A simple method to routinely monitor higher order structure changes and relation to
Michel Degueldre1, Annemie Wielant1, Eglantine Girot1
1Department of Analytical Science Biologicals, UCB , Braine L'Alleud , Belgium.
Native peptide mapping offers a straightforward method for analyzing higher order structure (HOS) in biopharmaceuticals. This technique proves effective for assessing the stability and structure-activity relationship of IgG antibodies, crucial for drug safety and efficacy.
Area of Science:
- Biopharmaceutical analysis
- Protein structure characterization
Background:
- Controlling Critical Quality Attributes (CQAs) like higher order structure (HOS) is vital for biopharmaceutical safety and efficacy.
- Traditional HOS analysis methods are often not suitable for routine quality control laboratories.
Purpose of the Study:
- To introduce a novel, quality control-amenable peptide mapping method for HOS analysis.
- To evaluate the suitability of native peptide mapping as a stability-indicating assay for IgG antibodies.
- To explore the relationship between HOS and biological activity of biotherapeutics.
Main Methods:
- Development and application of a native peptide mapping technique for HOS assessment.
- Utilizing heat stress to induce unfolding and studying the pathway via native peptide mapping.
- Correlating HOS changes (monitored by peptide mapping) with biological activity using cell-based assays.
Main Results:
- Native peptide mapping successfully identified structural differences between stressed and unstressed IgG4 and IgG1 antibodies, confirming its stability-indicating capability.
- The method elucidated the unfolding pathway of antibodies under heat stress.
- A significant correlation was observed between HOS alterations in complementary-determining regions and changes in relative potency.
Conclusions:
- Native peptide mapping is a fit-for-purpose assay for HOS analysis in quality control settings.
- This method effectively links higher order structure to biological activity, supporting its criticality as a CQA.
- The technique provides a valuable approach for interrogating the importance of HOS in biotherapeutic products.
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