Determination of influenza B identity and potency in quadrivalent inactivated influenza vaccines using lineage-specific monoclonal antibodies

Abstract

Co-circulation of two antigenically and genetically distinct lineages of influenza B virus, represented by prototype viruses B/Victoria/2/1987 and B/Yamagata/16/1988, has led to the development of quadrivalent influenza vaccines that contain two influenza B antigens. The inclusion of two influenza B antigens presents challenges for the production and regulation of inactivated quadrivalent vaccines, including the potential for cross-reactivity of the reagents used in identity and potency assays because of the relative close relatedness of the hemagglutinin (HA) from the two virus lineages. Monoclonal antibodies (mAbs) specific for the two lineages of influenza B HA were generated and characterized and used to set-up simple identity tests that distinguish the influenza B antigens in inactivated trivalent and quadrivalent vaccines. The lineage-specific mAbs bound well to the HA of influenza B strains included in influenza vaccines over a period of more than 10 years, suggesting that identity tests using such lineage-specific mAbs would not necessarily have to be updated with every influenza B vaccine strain change. These lineage-specific mAbs were also used in an antibody capture ELISA format to quantify HA in vaccine samples, including monovalent, trivalent, and quadrivalent vaccine samples from various manufacturers. The results demonstrated correlation with HA values determined by the traditional single radial immunodiffusion (SRID) assay. Further, the antibody-capture ELISA was able to distinguish heat-stressed vaccine from unstressed vaccine, and was similar to the SRID in quantifying the resultant loss of potency. These mAb reagents should be useful for further development of antibody-based alternative influenza B identity and potency assays.

Potency determination of inactivated H7 influenza vaccines using monoclonal antibody‐based ELISA and biolayer interferometry assays

“…There was generally good correlation between the SRID potency values and potency values obtained using either the mAb‐capture ELISA or BLI assay for most of the vaccine samples tested. However, as has been observed and discussed previously,9, 26 there are occasionally discrepancies in the actual values determined by SRID and any alternative potency assay, which may be at least partially related to the type of reference antigen used in the comparative analyses; additional work will be needed to better understand and resolve this issue. In the potency ELISA results reported here, one mAb (7B5) yielded potency results that were significantly lower for Vaccine 1 than the other tested mAbs.…”

Standardisation of inactivated influenza vaccines—Learning from history

“…This means that SRID assays can be used to assay each component of a trivalent vaccine without any interference. The recent development of quadrivalent vaccines containing antigens for the two lineages of influenza B has been a somewhat more challenging issue for SRID testing20 because there is some degree of cross‐reactivity between antibodies to the two lineages. It is nevertheless important that the standard antigen and the vaccine are antigenically homologous.…”

“…Universal antibodies that recognise all influenza subtypes have been described and used in an ELISA format to quantify HA, but these antibodies cannot distinguish influenza subtypes and may not be stability‐indicating because they bind HA under denaturing conditions 40. In other studies, strain‐specific mAbs have been used to quantify HA in capture ELISAs20, 41, 42; these assays are able to quantify each HA subtype antigen in multivalent vaccine formulations and are able to detect loss of potency in vaccine samples subjected to stress conditions. Another antibody‐binding platform under development as a possible potency assay uses panels of mAbs printed onto a slide format 43, 44.…”

“…However, these reagents may not be suitable for assays that measure properties of the vaccine antigen that are different from those measured by SRID. It has been suggested by several groups that one of the possible reasons why there is often correlation, but not always equivalence, between SRID potency results and potency values determined by alternative methods is that the current SRID antigen standards are whole‐inactivated influenza virus and are quite different from the vaccine formulation 20, 43. Some studies have addressed this issue by preparing a working standard that is similar in form to the vaccines being tested,41, 44 but many unanswered questions remain such as the actual extent and impact of the perceived problem, and whether assay‐specific or product‐specific standards are needed.…”

Development and use of Lentiviral Vectors Pseudotyped with Influenza B Haemagglutinins: application to vaccine immunogenicity, mAb potency and sero-surveillance studies

“…The seal serum samples (n=8) utilized were taken from seals caught in the Caspian Sea (CS) from 2006-2010, at the Europe/Asia border. Neutralisation of the full set of influenza B PV was evaluated using a panel of 5 previously characterised monoclonal antibodies from the Division of Viral Products, Food and Drug Administration, USA, see Table 1 (Verma 2017).…”