Characterization of a recombinant influenza vaccine candidate using complementary LC-MS methods

Area of Science:

• Biochemistry
• Analytical Chemistry
• Vaccinology

Background:

• Influenza vaccination is key to reducing illness and death from seasonal flu.
• Cell culture-based production offers advantages over traditional egg-based methods for influenza vaccines.
• Changes in viral antigens necessitate robust methods for vaccine characterization.

Purpose of the Study:

• To develop and apply advanced liquid chromatography-mass spectrometry (LC-MS) methods for characterizing cell-cultured influenza vaccines.
• To simultaneously identify hemagglutinin (HA) proteins and process-related impurities.
• To profile N-linked glycosylation sites and glycoforms on recombinant HA (rHA) antigens.

Main Methods:

• Reversed-phase LC-MS(E) for peptide mapping and characterization of rHA proteins.
• Hydrophilic interaction LC (HILIC)-multiple reaction monitoring (MRM) assays for glycoform confirmation and quantification.
• Utilized insect cell-baculovirus expression vector system (BEVS) for recombinant HA production.

Main Results:

• Successfully identified HA proteins and characterized site-specific N-linked glycosylation and degradation patterns.
• LC-MS(E) provided comprehensive protein identity and glycosylation data.
• HILIC-MRM enabled rapid confirmation and quantification of glycoforms and potential degradations.

Conclusions:

• Developed advanced LC-MS methods for comprehensive characterization of cell-cultured influenza vaccines.
• These methods are valuable for monitoring vaccine quality and supporting product comparability studies.
• Ensures the reliability and effectiveness of influenza vaccines produced via cell culture.