Anti-P. falciparum AMA1 Recombinant Antibody (MRO-191CT) (CAT#: MRO-191CT)

Figure 1 V(NAR)s 12Y-1 and 12Y-2 are specific for P. falciparum AMA1.

ELISA comparison of binding of affinity purified proteins 12Y-1 (black) and 12Y-2 (grey) to lysozyme, AMA1, -amylase, and Tom70.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

Figure 2 ELISA of V(NAR) 12Y-2 by error-prone PCR.

After three rounds of panning, 16 affinity-matured clones were tested for binding to AMA1 and to lysozyme (negative control antigen). The hatched line represents the parental (12Y-2) response level.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

Figure 3 The 12Y-2 triple-mutant shows parental affinity for AMA1.

BIAcore sensorgrams comparing the binding of identical concentrations of the parental (12Y-2), single mutant (14M-15), and triple mutant proteins to immobilized AMA1.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

Figure 4 Binding was measured at a constant flow rate of 5 ul/min with an injection volume of 35 ul. Dissociation was continued with HBS buffer until the response returned to the initial value before injecting the next sample

V(NAR)s 12Y-1 and 12Y-2 are specific for P. falciparum AMA1.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

Figure 5 Binding was measured at a constant flow rate of 5 ul/min with an injection volume of 35 ul. Dissociation was continued with HBS buffer until the response returned to the initial value before injecting the next sample

V(NAR)s 12Y-1 and 12Y-2 are specific for P. falciparum AMA1.

Nuttall, S. D., Humberstone, K. S., Krishnan, U. V., Carmichael, J. A., Doughty, L., Hattarki, M., ... & Irving, R. A. (2004). Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1. PROTEINS: Structure, Function, and Bioinformatics, 55(1), 187-197.

Figure 6 IgNAR constant domain 1 is insufficient for dimer formation.

BIAcore sensorgrams comparing binding of proteins 17T-6 and 12Y-2 to immobilised AMA1 protein.

Simmons, D. P., Abregu, F. A., Krishnan, U. V., Proll, D. F., Streltsov, V. A., Doughty, L., ... & Nuttall, S. D. (2006). Dimerisation strategies for shark IgNAR single domain antibody fragments. Journal of immunological methods, 315(1-2), 171-184.

Figure 7 Dhlx-mediated dimerisation.

The peak-purified dimer fraction was analysed by BIAcore biosensor, compared to the parental 12Y-2

Simmons, D. P., Abregu, F. A., Krishnan, U. V., Proll, D. F., Streltsov, V. A., Doughty, L., ... & Nuttall, S. D. (2006). Dimerisation strategies for shark IgNAR single domain antibody fragments. Journal of immunological methods, 315(1-2), 171-184.

Figure 8 Tandem V(NAR)s in a single chain format

BIAcore sensorgrams comparing binding of proteins 12Y-2, 14M-15, and 15Q-1 to immobilised AMA1 protein.

Simmons, D. P., Abregu, F. A., Krishnan, U. V., Proll, D. F., Streltsov, V. A., Doughty, L., ... & Nuttall, S. D. (2006). Dimerisation strategies for shark IgNAR single domain antibody fragments. Journal of immunological methods, 315(1-2), 171-184.

Figure 9 ELISA analysis of 40 12Y-2 variants binding to AMA-1 and a control negative antigen.

ELISA analysis of 40 12Y-2 variants binding to AMA-1 and a control negative antigen.

Figure 10 Parasite Invasion Inhibition

Recombinant V(NAR) domains inhibit invasion of erythrocytes by P. falciparum 3D7 parasites. Data are represented as the mean ± standard error of quadruplicate experiments.

Henderson, K. A., Streltsov, V. A., Coley, A. M., Dolezal, O., Hudson, P. J., Batchelor, A. H., ... & Foley, M. (2007). Structure of an IgNAR-AMA1 complex: targeting a conserved hydrophobic cleft broadens malarial strain recognition. Structure, 15(11), 1452-1466.

Figure 11 Parasite Invasion Inhibition

Recombinant V(NAR) domains inhibit invasion of erythrocytes by P. falciparum 3D7 parasites. Data are represented as the mean ± standard error of quadruplicate experiments,except for P. falciparum W2mef parasites.

Henderson, K. A., Streltsov, V. A., Coley, A. M., Dolezal, O., Hudson, P. J., Batchelor, A. H., ... & Foley, M. (2007). Structure of an IgNAR-AMA1 complex: targeting a conserved hydrophobic cleft broadens malarial strain recognition. Structure, 15(11), 1452-1466.