tetano
Editor, Senior Moderator
Appl Microbiol Biotechnol
. 2021 May 3.
doi: 10.1007/s00253-021-11247-5. Online ahead of print.
Comparison of N-linked glycosylation on hemagglutinins derived from chicken embryos and MDCK cells: a case of the production of a trivalent seasonal influenza vaccine
Jingqi Li[SUP] 1 [/SUP], Sixu Liu[SUP] 1 [/SUP], Yanlin Gao[SUP] 1 2 [/SUP], Shuaishuai Tian[SUP] 1 [/SUP], Yu Yang[SUP] 3 [/SUP], Ningning Ma[SUP] 4 [/SUP]
Affiliations
Abstract
N-linked glycosylation plays critical roles in folding, receptor binding, and immunomodulating of hemagglutinin (HA), the main antigen in influenza vaccines. Chicken embryos are the predominant production host for influenza vaccines, but Madin-Darby canine kidney (MDCK) cells have emerged as an important alternative host. In this study, we compared glycosylation patterns, including the occupancy of potential glycosylation sites and the distribution of different glycans, on the HAs of three strains of influenza viruses for the production a trivalent seasonal flu vaccine for the 2015-2016 Northern Hemisphere season (i.e., A/California/7/2009 (H1N1) X179A, A/Switzerland/9715293/2013 (H3N2) NIB-88, and B/Brisbane/60/2008 NYMC BX-35###). Of the 8, 12, and 11 potential glycosylation sites on the HAs of H1N1, H3N2, and B strains, respectively, most were highly occupied. For the H3N2 and B strains, MDCK-derived HAs contained more sites being partially occupied (<95%) than embryo-derived HAs. A highly sensitive glycan assay was developed where 50 different glycans were identified, which was more than what has been reported previously, and their relative abundance was quantified. In general, MDCK-derived HAs contain more glycans of higher molecular weight. High-mannose species account for the most abundant group of glycans, but at a lower level as compared to those reported in previous studies, presumably due to that lower abundance, complex structure glycans were accounted for in this study. The different glycosylation patterns between MDCK- and chicken embryo-derived HAs may help elucidate the role of glycosylation on the function of influenza vaccines. KEY POINTS: ? For the H3N2 and B strains, MDCK-derived HAs contained more partially (<95%) occupied glycosylation sites. ? MDCK-derived HAs contained more glycans of higher molecular weight. ? A systematic comparison of glycosylation on HAs used for trivalent seasonal flu vaccines was conducted.
Keywords: Glycan occupancy; Hemagglutinin; Influenza vaccine; Madin-Darby canine kidney (MDCK) cells; N-linked glycosylation.
. 2021 May 3.
doi: 10.1007/s00253-021-11247-5. Online ahead of print.
Comparison of N-linked glycosylation on hemagglutinins derived from chicken embryos and MDCK cells: a case of the production of a trivalent seasonal influenza vaccine
Jingqi Li[SUP] 1 [/SUP], Sixu Liu[SUP] 1 [/SUP], Yanlin Gao[SUP] 1 2 [/SUP], Shuaishuai Tian[SUP] 1 [/SUP], Yu Yang[SUP] 3 [/SUP], Ningning Ma[SUP] 4 [/SUP]
Affiliations
- PMID: 33937925
- DOI: 10.1007/s00253-021-11247-5
Abstract
N-linked glycosylation plays critical roles in folding, receptor binding, and immunomodulating of hemagglutinin (HA), the main antigen in influenza vaccines. Chicken embryos are the predominant production host for influenza vaccines, but Madin-Darby canine kidney (MDCK) cells have emerged as an important alternative host. In this study, we compared glycosylation patterns, including the occupancy of potential glycosylation sites and the distribution of different glycans, on the HAs of three strains of influenza viruses for the production a trivalent seasonal flu vaccine for the 2015-2016 Northern Hemisphere season (i.e., A/California/7/2009 (H1N1) X179A, A/Switzerland/9715293/2013 (H3N2) NIB-88, and B/Brisbane/60/2008 NYMC BX-35###). Of the 8, 12, and 11 potential glycosylation sites on the HAs of H1N1, H3N2, and B strains, respectively, most were highly occupied. For the H3N2 and B strains, MDCK-derived HAs contained more sites being partially occupied (<95%) than embryo-derived HAs. A highly sensitive glycan assay was developed where 50 different glycans were identified, which was more than what has been reported previously, and their relative abundance was quantified. In general, MDCK-derived HAs contain more glycans of higher molecular weight. High-mannose species account for the most abundant group of glycans, but at a lower level as compared to those reported in previous studies, presumably due to that lower abundance, complex structure glycans were accounted for in this study. The different glycosylation patterns between MDCK- and chicken embryo-derived HAs may help elucidate the role of glycosylation on the function of influenza vaccines. KEY POINTS: ? For the H3N2 and B strains, MDCK-derived HAs contained more partially (<95%) occupied glycosylation sites. ? MDCK-derived HAs contained more glycans of higher molecular weight. ? A systematic comparison of glycosylation on HAs used for trivalent seasonal flu vaccines was conducted.
Keywords: Glycan occupancy; Hemagglutinin; Influenza vaccine; Madin-Darby canine kidney (MDCK) cells; N-linked glycosylation.