Fig. 6

Purity of phage display light chain shuffled antibodies as Immunoglobulin G1 and indirect ELISA against AMA1-variants. Antibodies were transiently expressed as human IgG1/κ in N. benthamiana and purified using Protein A-chromatography. a SDS-PAGE Coomassie Brilliant blue stained, b Western blot, anti-human-IgG (H+L)-AP (1:5000) + NBT/BCIP. Dividing lines in B indicate parts from the same blot that were re-arranged. c Indirect ELISA against AMA1-variants with plant-expressed human IgG1/κ. The four selected light chain-shuffled antibodies OcheA-9, OcheA-16, OcheA-17 and OcheA-17B, parental antibody OcheA and AMA1-specific antibodies 1E4 and 4G2 were analysed in the presence (100 ng/well) and absence (PBS) of the antigens DiCo1-3 and AMA1-GKO (glyco-knockout, strain 3D7). Values represent mean ± SD of triplicate measurements after subtraction of the secondary antibody-control values