Recombinant Mouse Anti-P. vivax AMA-1 Antibody (F8.12.19) (CAT#: PABZ-098)

Figure 1 Surface plasmon resonance runs showing the interaction of the recombinant ectoplasmic region of AMA1 as analyte and IgG F8.12.19 covalently immobilised on the Biacore sensor chip.

The runs were corrected for non-specific binding by subtraction of the curves obtained by passage of the same protein solution through a blank channel (no immobilised IgG) on the same senor chip. The curves labelled buffer show the results for passage of the running buffer alone. Analyte was injected for 60 s for the association phase. This was followed by injection of the running buffer alone at the same flow-rate to give the dissociation phase. The response in resonance units (RU) is plotted as a function of time (in s.). (a) PvAMA1 at concentrations 5.0 μg/ml, 2.5 μg/ml, 1.25 μg/ml and 0.62 μg/ml, as indicated. (b) PfAMA1 at concentrations 50 μg/ml, 25 μg/ml, 12.5 μg/ml and 6.2 μg/ml, as indicated.

Igonet, S., Vulliez-Le Normand, B., Faure, G., Riottot, M. M., Kocken, C. H., Thomas, A. W., & Bentley, G. A. (2007). Cross-reactivity studies of an anti-Plasmodium vivax apical membrane antigen 1 monoclonal antibody: binding and structural characterisation. Journal of molecular biology, 366(5), 1523-1537.

Figure 2 RON2-AMA1 interaction is conserved in P. falciparum.

(A) Pf3D7-AMA1 displays a type I TM topology at the plasma membrane of BHK-21 cells. PfAMA1 transfected BHK-21 cells were subjected to IFA. Mouse MAb F8.12.19 recognizes extracellular PfAMA1 domain III that is accessible with or without permeabilization, whereas anti-Myc antibody labels the intracellular C-terminal Myc tag in permeabilized cells only. (B) PfAMA1 transfected BHK-21 cells were incubated with 20 µg/ml of GST or PfRON2-5. Recombinant proteins were detected using anti-GST antibody.

Lamarque, M., Besteiro, S., Papoin, J., Roques, M., Vulliez-Le Normand, B., Morlon-Guyot, J., ... & Kocken, C. H. (2011). The RON2-AMA1 interaction is a critical step in moving junction-dependent invasion by apicomplexan parasites. PLoS pathogens, 7(2), e1001276.

Figure 3 SDS‐PAGE and immunoblot blot analysis.

(c) SDS‐PAGE analysis of purified PfAMA1 variants under reducing and nonreducing conditions and (d) corresponding immunoblot under nonreducing conditions. The PfAMA1 variants migrate as expected with 60.4 kDa for PPkoAMA1 including the 8.7‐kDa propeptide sequence (lane 1), 53.6 kDa for koAMA1 (lane 2) and 58.4 kDa for gAMA1 (lane 3), respectively. Proteolytic clipping was observed, as previously described, for PPkoAMA1 resulting in additional bands of 55 kDa and 27 kDa under reducing conditions. Identity of the PfAMA1 was confirmed by the binding of PfAMA1‐specific antibodies (1F9, 4G2, F8.12.19) in the immunoblot.

Boes, A., Spiegel, H., Edgue, G., Kapelski, S., Scheuermayer, M., Fendel, R., ... & Pradel, G. (2015). Detailed functional characterization of glycosylated and nonglycosylated variants of malaria vaccine candidate Pf AMA 1 produced in Nicotiana benthamiana and analysis of growth inhibitory responses in rabbits. Plant biotechnology journal, 13(2), 222-234.