Development and validation of a peptide-mapping procedure for a novel C1 esterase inhibitor

Peptide mapping is a key method for studying the primary structure of proteins. With its sensitivity to slightest changes in covalent protein, this applicable both medicinal product identification at control stage and production process stability monitoring. The aim study was develop validate peptide-mapping procedure novel highly glycosylated recombinant C1 esterase inhibitor. Materials methods. authors studied human inhibitor products trypsin. involved peptide using reverse-phase high-performance liquid chromatography high-resolution mass spectrometry. following statistics were calculated evaluate results: mean, standard deviation, coefficient variation. validation parameters included specificity, precision, robustness. Results. tested several variants sample preparation tryptic digests, including additional N-glycanase treatment complete deglycosylation, established optimal conditions chromatographic separation peptides obtain consistent profiles (peptide maps). identified characteristic peaks corresponding relative retention time area ranges. absolute second (characteristic) peak approximately 16.5–16.9 minutes. times 2.14–2.21 9, 2.55–2.64 12, 2.97–3.14 14, 3.11–3.29 15, 6.20–6.63 28. Conclusions. developed inhibitors optimised achieve an over 18-hour reduction time. This met acceptance criteria