Table 3. Antibody classification.
Clone | Antibody neutralisation category1 | HI titre (µg/ml) H1N12 | H1 (D18-I530)3 KD (nM) | HA1 (H1N1) (D18-R344)3 KD (nM) | HA4 P66-I282 | HA4 L193-K225 | HA4 D363-G478 | Low pH treated HA 5 | Neutral pH treated HA 5 |
R1a-G5 | R | 1.56 | 5.85 | 15.6 | + | - | - | + | + |
R2b-D8 | N | - | - | - | - | - | + | ++ | + |
R2b-E8 | C | - | 1.74 | - | - | - | - | - | + |
R2b-D9 | C | - | 0.18 | - | - | - | - | - | + |
R1a-F4 | R | 2.08 | 6.46 | 4.74 | + | - | - | + | + |
R1a-A5 | C | - | 1.87 | - | - | - | - | - | + |
R1a-C5 | R | 10.40 | 37.57 | 38.40 | + | + | - | + | + |
R1a-E5 | R | - | 0.79 | - | - | - | - | - | + |
R1a-F5 | N | - | 10.00 | 6.06 | + | - | - | + | + |
R1a-B6 | C | - | 0.86 | - | - | - | - | - | + |
R1a-E6 | N | - | 9.97 | 36.00 | + | - | - | + | + |
R1a-G6 | R | 1.04 | 4.83 | 4.62 | + | - | - | + | + |
R2a-E8 | R | - | 0.79 | - | - | - | - | - | + |
R2a-G8 | C | - | 3.55 | 7.66 | - | - | - | - | + |
R2a-B9 | R | - | 0.16 | - | - | - | - | - | + |
R2a-F9 | R | 1.04 | 78.80 | 78.80 | + | - | - | + | + |
R2a-G9 | R | 1.30 | 22.14 | 68.70 | + | - | - | + | + |
R2a-H9 | R | 1.30 | 2.77 | 468.00 | + | - | - | + | + |
control | - | - | - | - | - | - | - | - | - |
Antibody neutralisation category, R is H1N1 restricted neutralisation activity, C refers to a cross-neutralising antibody, N is non-neutralising antibody
Hemagglutination inhibition (HI) titre was given as the minimum dilution of antibody at which inhibition of agglutination of turkey erythrocytes was observed (in µg/ml of purified antibody). Values are given as an average of three independent assays. Laboratory adapted A(H1N1) strain (X-181) was used.
Single cycle affinity determination on full length HA (D18-I530) and globular head HA1 domain (residues D18-R344) derived from A/California/06/2009 (H1N1pdm09). Affinity is given in nM.
Binding to phage displayed fragments of HA as indicated in ELISA (see Figure 5B).
ELISA on HA of A/California/07/2009 (H1N1pdm09) treated with low pH and neutral pH (see Figure 5A).