pr400329k_si_001.pdf (865.13 kB)
Comparative Glycomics Analysis of Influenza Hemagglutinin (H5N1) Produced in Vaccine Relevant Cell Platforms
journal contribution
posted on 2013-08-02, 00:00 authored by Yanming An, Joseph A. Rininger, Donald
L. Jarvis, Xianghong Jing, Zhiping Ye, Jared J. Aumiller, Maryna Eichelberger, John F. CipolloHemagglutinin (HA)
is the major antigen in influenza vaccines,
and glycosylation is known to influence its antigenicity. Embryonated
hen eggs are traditionally used for influenza vaccine production,
but vaccines produced in mammalian and insect cells were recently
licensed. This raises the concern that vaccines produced with different
cell systems might not be equivalent due to differences in their glycosylation
patterns. Thus, we developed an analytical method to monitor vaccine
glycosylation through a combination of nanoLC/MSE and quantitative
MALDI-TOF MS permethylation profiling. We then used this method to
examine glycosylation of HAs from two different influenza H5N1 strains
produced in five different platforms, including hen eggs, three different
insect cell lines (High Five, expresSF+ and glycoengineered expresSF+), and a human cell line (HEK293). Our results
demonstrated that (1) sequon utilization is not necessarily equivalent
in different cell types, (2) there are quantitative and qualitative
differences in the overall N-glycosylation patterns
and structures produced by different cell types, (3) ∼20% of
the N-glycans on the HAs produced by High Five cells
are core α1,3-fucosylated structures, which may be allergenic
in humans, and (4) our method can be used to monitor differences in
glycosylation during the cellular glycoengineering stages of vaccine
development.