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Poster Presentations
Day 2, June 23（Mon.）
Room P (Maesato East, Foyer, Ocean Wing)
- 2P-PM-07
Case study of inter-laboratory multi-attribute method (MAM) data comparison in Japan using NIST mAb
(Protein Metrics)
oAyako Kurimoto, Kadir Sen
Multi-attribute method (MAM), a liquid chromatography-mass spectrometry (LC-MS)-based peptide mapping method, has become widely used in protein therapeutics industry in recent years. In a round-robin study in Japan, participants performed LC-MS analysis of the same lot of tryptic digestion and undigested commercial NIST mAb. The purpose of this inter-laboratory MAM study was to confirm and refine the basic techniques used by each laboratory to perform MAM of antibody therapeutics.
Purchased NIST mAb Tryptic Digestion Standard (purchased digested sample) and NIST humanized IgG1κ monoclonal antibody (native sample) were distributed to the participants, and the native sample was subjected to tryptic digestion in each laboratory according to a previously reported method1). The collected raw data were processed in Byosphere based on the same fifteen reference peptides as the MAM Consortium Round Robin study2).
We compared the relative abundance of several predefined reference NIST mAb CQAs (deamidation, ammonia-loss, oxidation, terminal Lys-loss, and pyroQ). The sum of the percent mod for the oxidation of peptide DTLMISR had some outliers in two labs data. For the deamidated peptide a PENNY peptide, the sum of the percent mod did not show any outliers.
References
1)S.Millán-Martín et al., Nat Protoc., 18(4), 1056-1089 (2023).
2)T.Mouchahoir et al., J. Am. Soc. Mass Spectrom., 33, 1659-1677 (2022).