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Vaccine . Bromelain-cleaved hemagglutinin production from cell culture-derived influenza viruses enhances vaccine potency quantification by single r

tetano

Editor, Senior Moderator
Vaccine


. 2026 Jan 16:75:128235.
doi: 10.1016/j.vaccine.2026.128235. Online ahead of print.
Bromelain-cleaved hemagglutinin production from cell culture-derived influenza viruses enhances vaccine potency quantification by single radial immunodiffusion assay

Ohseok Jeong[SUP] 1 [/SUP], Wooyoung Choi[SUP] 2 [/SUP], Hyebin Ahn[SUP] 3 [/SUP], Seonghyun Lee[SUP] 4 [/SUP], Yong Wook Park[SUP] 5 [/SUP], Hun Kim[SUP] 6 [/SUP], Jae-Hwan Nam[SUP] 7 [/SUP]


Affiliations
Abstract

Single Radial Immunodiffusion (SRID) assay remains the gold standard for determining influenza vaccine potency. Accurate SRID assays require high-purity antigens and strain-specific antisera that closely match circulating viruses. However, mutations in influenza viruses propagated in traditional egg-based systems affect antigenicity and assay accuracy. Therefore, we optimized bromelain-cleaved hemagglutinin (BHA) production from cell culture-derived influenza viruses, specifically targeting A/H3N2 and B/Victoria lineage strains. We employed a sequential enzymatic digestion strategy and successfully generated highly purified BHA with improved antigen yield and minimal neuraminidase contamination. The resulting antisera demonstrated strong, strain-specific reactivity. Subsequent SRID assays confirmed that homologous antigen-antiserum pairs derived from cell culture-derived materials provided significantly more accurate hemagglutinin quantification than the heterologous or egg-derived combinations. These findings highlight the need to match antigen and antiserum sources. Further, cell culture-derived reagents could be used to enhance assay reliability, advancing influenza vaccine standardization and quality control in cell-based vaccine production. By demonstrating the importance of antigen-antiserum matching and optimizing BHA production from cell culture-derived influenza viruses, this study establishes a practical foundation for improving the reliability of SRID-based potency testing and advancing the standardization of next-generation influenza vaccines.

Keywords: Antigen standardization; Bromelain; Cell culture; Influ

 
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