Characterization of PEGylation sites in Neulasta and a biosimilar candidate with a combined fragmentation strategy in mass spectrometry analysis.

Autor: Rauniyar N; Tanvex BioPharma USA, Inc., San Diego, California, USA., Togle AJ; Tanvex BioPharma USA, Inc., San Diego, California, USA., Ronci RA; Tanvex BioPharma USA, Inc., San Diego, California, USA., Reyes D; Tanvex BioPharma USA, Inc., San Diego, California, USA., Han X; Tanvex BioPharma USA, Inc., San Diego, California, USA.
Jazyk: angličtina
Zdroj: Journal of mass spectrometry : JMS [J Mass Spectrom] 2024 Apr; Vol. 59 (4), pp. e5017.
DOI: 10.1002/jms.5017
Abstrakt: In the development of biosimilar products to Neulasta, it is essential to determine the intact molecular mass and confirm precise PEGylation sites. In this study, we applied a combination of techniques, including post-column addition of triethylamine in reversed-phase liquid chromatography-mass spectrometry (RPLC-MS) to determine the intact molecular mass, and in-source fragmentation (ISF) and higher-energy collision dissociation-tandem mass spectrometry (HCD-MS/MS) to identify the PEGylation site. Our results show that both the pegfilgrastim biosimilar candidate and Neulasta lots are mono-PEGylated at the N-terminal end. Furthermore, we show that the combined ISF and HCD-MS/MS method can be used for identifying the PEGylation sites in the diPEGylated variant of pegfilgrastim. The diPEGylated variant has modification sites at the N-terminal end and a lysine at position 35 in the protein sequence.
(© 2024 John Wiley & Sons Ltd.)
Databáze: MEDLINE
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