Table 1.
Influenza B HA mAbs.
Fig 1.
Location of HA amino acid changes in influenza B escape mutants.
(A) Antigenic structure of the B/Victoria lineage HA trimer (B/Brisbane/60/2008 PDB ID: 4FQM) and location of the escape mutants to mAbs BR5A1 (blue), BR8E12 (magenta), BR7B7 (yellow), and CR2F11 (green). (B) Antigenic structure of the B/Yamagata lineage HA trimer (B/Yamanashi/166/1998 PDB ID: 4M40) and location of the escape mutants to mAbs MA1H4 (orange), WI3E8 (red), MA3B2 (cyan), and CR2F11 (green). Escape mutants to the cross-reactive mAb CR2F11 were derived from both B/Victoria and B/Yamagata viruses.
Table 2.
mAb neutralization of B/Victoria escape mutants.
Table 3.
mAb neutralization of B/Yamagata escape mutants.
Fig 2.
SRID analysis of quadrivalent influenza vaccines using sheep polyclonal antiserum produced to B/Brisbane/60/2008.
Dilutions of quadrivalent influenza vaccine containing B/Brisbane/60/2008 and B/Massachusetts/2/2012 (A) or B/Brisbane/60/2008 and B/Texas/02/2013 (B) were loaded onto agarose gels (rows 2) along with dilutions of the two corresponding reference antigens (rows 1 and 3) and analyzed by standard SRID using B/Brisbane/60/2008 reference antiserum (Lot B-Ab-1108).
Fig 3.
ELISA identity analysis of trivalent and quadrivalent influenza vaccines using lineage-specific mAbs.
ELISA plates were coated with the indicated purified mAbs at 2 μg/ml and used to capture Reference Antigens B/Brisbane/60/2008 (60 μg/ml) and B/Massachusetts/2/2012 (58 μg/ml), a quadrivalent vaccine containing B/Brisbane and B/Mass antigens at 24 and 40 μg/ml, respectively, a trivalent vaccine containing B/Florida/4/2006 (25 μg/ml), and a trivalent vaccine containing B/Brisbane (27 μg/ml). The starting dilution for all reference antigens and vaccines was 1:300.
Fig 4.
Lineage specificity of mAb-capture ELISA.
ELISA plates were coated with the indicated purified mAbs at 2 μg/ml and used to capture Reference Antigens B/Brisbane/60/2008 (60 μg/ml), B/Malaysia/2506/2004 (76 μg/ml), B/Hong Kong/330/2001 (69 μg/ml), B/Massachusetts/2/2012 (58 μg/ml), B/Florida/4/2006 (79 μg/ml), B/Texas/6/2011 (80 μg/ml), and B/Phuket/3073/2013 (78 μg/ml). The starting dilution for all reference antigens was 1:300.
Fig 5.
Correlation between SRID potency values and ELISA potency values determined for vaccines containing B/Victoria antigens.
(A) Eight vaccine samples, including 3 monovalent vaccines (■), 2 trivalent vaccines (▲), and 3 quadrivalent vaccines (●) were analyzed for HA content by standard SRID and mAb-capture ELISA using B/Victoria mAbs BR5A1 (closed symbols) and BR8E12 (open symbols). (B) SRID potency values for each vaccine plotted against the mAb-capture ELISA potency value using the combined (mean) BR5A1 and BR8E12 ELISA potency values.
Fig 6.
Correlation between SRID potency values and ELISA potency values determined for vaccines containing B/Yamagata antigens.
(A) Six vaccine samples, including 3 trivalent vaccines (▲) and 3 quadrivalent vaccines (●) were analyzed for HA content by standard SRID and mAb-capture ELISA using B/Yamagata mAbs MA1H4 (closed symbols) and WI3E8 (open symbols). (B) SRID potency values for each vaccine plotted against the mAb-capture ELISA potency value using the combined (mean) MA1H4 and WI3E8 ELISA potency values.
Table 4.
SRID and ELISA influenza B potency of influenza quadrivalent vaccine subjected to temperature stress at 56°C.