tetano
Editor, Senior Moderator
Vaccine
. 2025 Apr 21:56:127157.
doi: 10.1016/j.vaccine.2025.127157. Online ahead of print. Fluorescence-barcoded cell lines stably expressing membrane-anchored influenza neuraminidases
Joel Finney[SUP] 1 [/SUP], Masayuki Kuraoka[SUP] 2 [/SUP], Shengli Song[SUP] 3 [/SUP], Akiko Watanabe[SUP] 2 [/SUP], Xiaoe Liang[SUP] 2 [/SUP], Dongmei Liao[SUP] 2 [/SUP], M Anthony Moody[SUP] 4 [/SUP], Emmanuel B Walter[SUP] 5 [/SUP], Stephen C Harrison[SUP] 6 [/SUP], Garnett Kelsoe[SUP] 7 [/SUP]
Affiliations
The discovery of broadly protective antibodies to the influenza virus neuraminidase (NA) has raised interest in NA as a vaccine target. However, recombinant, solubilized tetrameric NA ectodomains are often challenging to express and isolate, hindering the study of anti-NA humoral responses. To address this obstacle, we established a panel of 22 non-adherent cell lines stably expressing native, historical N1, N2, N3, N9, and NB NAs anchored on the cell surface. The cell lines are barcoded with fluorescent proteins, enabling high-throughput, 16-plex analyses of antibody binding with commonly available flow cytometers. The cell lines were at least as efficient as a Luminex multiplex binding assay at identifying NA antibodies from a library of unselected clonal IgGs derived from human memory B cells. The cell lines were also useful for measuring the magnitude and breadth of the serum antibody response elicited by experimental infection of rhesus macaques with influenza virus. The membrane-anchored NAs are catalytically active and are compatible with established sialidase activity assays. NA-expressing K530 cell lines therefore represent a useful tool for studying NA immunity and evaluating influenza vaccine efficacy.
Keywords: Barcode; Barcoding; Binding assays; ELLA; Flow cytometry; Fluorescent protein; Immunoassay; NA-Star assay; Sialidase; Vaccine.
. 2025 Apr 21:56:127157.
doi: 10.1016/j.vaccine.2025.127157. Online ahead of print. Fluorescence-barcoded cell lines stably expressing membrane-anchored influenza neuraminidases
Joel Finney[SUP] 1 [/SUP], Masayuki Kuraoka[SUP] 2 [/SUP], Shengli Song[SUP] 3 [/SUP], Akiko Watanabe[SUP] 2 [/SUP], Xiaoe Liang[SUP] 2 [/SUP], Dongmei Liao[SUP] 2 [/SUP], M Anthony Moody[SUP] 4 [/SUP], Emmanuel B Walter[SUP] 5 [/SUP], Stephen C Harrison[SUP] 6 [/SUP], Garnett Kelsoe[SUP] 7 [/SUP]
Affiliations
- PMID: 40262372
- DOI: 10.1016/j.vaccine.2025.127157
The discovery of broadly protective antibodies to the influenza virus neuraminidase (NA) has raised interest in NA as a vaccine target. However, recombinant, solubilized tetrameric NA ectodomains are often challenging to express and isolate, hindering the study of anti-NA humoral responses. To address this obstacle, we established a panel of 22 non-adherent cell lines stably expressing native, historical N1, N2, N3, N9, and NB NAs anchored on the cell surface. The cell lines are barcoded with fluorescent proteins, enabling high-throughput, 16-plex analyses of antibody binding with commonly available flow cytometers. The cell lines were at least as efficient as a Luminex multiplex binding assay at identifying NA antibodies from a library of unselected clonal IgGs derived from human memory B cells. The cell lines were also useful for measuring the magnitude and breadth of the serum antibody response elicited by experimental infection of rhesus macaques with influenza virus. The membrane-anchored NAs are catalytically active and are compatible with established sialidase activity assays. NA-expressing K530 cell lines therefore represent a useful tool for studying NA immunity and evaluating influenza vaccine efficacy.
Keywords: Barcode; Barcoding; Binding assays; ELLA; Flow cytometry; Fluorescent protein; Immunoassay; NA-Star assay; Sialidase; Vaccine.