NIH Research Festival
The routinely used method to quantitate the haemagglutinin (HA) content in the influenza vaccines is Single Radial Immunodiffusion (SRID). SRID is not particularly sensitive; it is relatively low throughput and requires development of strain specific reagents which at times can be troublesome. The 2009 H1N1 pandemic highlighted the need to develop alternate assays to quickly and accurately quantitate the HA concentration in the vaccines and expedite the release of vaccine lots. We have developed an ELISA based assay for rapid influenza HA quantitation in the vaccine lots. Monoclonal antibodies (mAbs) and polyclonal sera specific to B/Brisbane/60/2008, B/Wisconsin/1/2010 and B/Massachusetts/2/2012 were produced by immunizing the virus like particles (VLPs) and plasmid DNA expressing the respective HA. The mAbs were characterized in ELISA, hemagglutination inhibition, western and neutralization assays. Selected mAbs were evaluated for their potential to capture the HA from reference antigen and different influenza vaccines. Vaccine samples included monovalent, trivalent and quadrivalent vaccine lots from different manufacturers. Generation of mAbs and antisera by immunizing VLPs and plasmid DNA expressing the strain-specific HA could shorter the timeline for the potency reagents preparation by couple of months. The quantitation of HA by ELISA based method correlated with the SRID data. The mAbs in ELISA did not show any cross reactivity as seen in some SRID sheep sera and therefore was used to perform vaccine identity testing. The mAb-capture ELISA is sensitive and accurate and has the potential to be used as an alternative to SRID during a pandemic.
Scientific Focus Area: Virology
This page was last updated on Friday, March 26, 2021