Anti-Influenza A Virus H1N1 Hemagglutinin HA1 subunit Recombinant Antibody (MRO-235LC-VHH) (CAT#: MRO-235LC-VHH)

Figure 1 Specificity of single domain antibodies to different influenza antigen reference reagents.

ELISA comparing binding of purified VHH antibodies at 30 mg/ml against H1N1, seasonal H1N1 and H5N1.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

Figure 2 Antibody affinity on recombinant HA by surface plasmon resonance.

Affinity on recombinant H1-HA, H1N1.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

Figure 3 Characterisation of antibody epitopes.

ELISA showing reactivity of purified antibodies at 30 mg/ml to HA antigen standard H1N1 either treated with low pH or neutral pH.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

Figure 4 Characterisation of antibody epitopes.

ELISA showing binding of phage displayed HA gene fragments to purified sdAbs.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

Figure 5 Grouping of antibody epitopes.

SPR co-injection experiments were used to determine if pairs of VHH antibodies can bind to recombinant H1-HA simultaneously. The antibodies R1a-A5, R1a-B6, R2b-D9, R2b-E8, R1a-C5 and R2a-G9 were tested in combination with each other. Example series of sensorgrams of R2b-D9 injected as the first antibody followed by injection of each of the other antibodies as indicated R2b-D9/R1a-C5.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.

Figure 6 Grouping of antibody epitopes.

The sensorgrams indicate the Rmax value for each of the individual antibodies with no significant increase in Rmax following injection of the antibody mixture, which suggests these antibodies recognise an overlapping epitope or an epitope that hinders binding of a second antibody. If the antibodies recognised non-overlapping or non-hindering sites the Rmax would be expected to be the sum of the individual Rmax values. This was confirmed when R1a-C5 which was predicted to recognise a non-overlapping epitope was included in a equimolar mixture of five antibodies and the response was seen to increase by an amount equivalent to R1a-C5 binding individually.

Hufton, S. E., Risley, P., Ball, C. R., Major, D., Engelhardt, O. G., & Poole, S. (2014). The breadth of cross sub-type neutralisation activity of a single domain antibody to influenza hemagglutinin can be increased by antibody valency. PLoS One, 9(8), e103294.