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Fig. 1 | Malaria Journal

Fig. 1

From: Analysis of the dose-dependent stage-specific in vitro efficacy of a multi-stage malaria vaccine candidate cocktail

Fig. 1

Antigen design, LDS-PAGE and immunoblot blot analysis of VAMAX 1, VAMAX 2 and VAMAX 6 expression. a Schematic representation of the VAMAX 1, VAMAX 2 and VAMAX 6 fusion antigens. DiCo1–3 diversity covering variants of PfAMA1 (red); Pfs25 P. falciparum 3D7 sexual-stage surface antigen Pfs25 (blue); PfCSP_TSR thrombospondin-related region from P. falciparum 3D7 circumsporozoite protein PfCSP (gray); PfMSP1_19 C-terminal 19 kDa fragment of merozoite surface protein 1 (PfMSP1_19) from P. falciparum strain FUP (pink); PfRH5_Q5A Short amino acid stretch of the reticulocyte-binding protein homologue 5 (PfRH5) of P. falciparum 3D7, epitope of the invasion inhibitory antibody mAb Q5A. b LDS-PAGE analysis under non-reducing conditions of VAMAX 1 (V1), VAMAX 2 (V2) and VAMAX6 (V 6) after purification (3.4 µg of each VAMAX fusion protein per lane). c Immunoblot analysis under non-reducing conditions of purified VAMAX fusion proteins (V1, V2 and V6): 1.3 µg of each VAMAX fusion protein was loaded and the fusion proteins (V1, V2 and V6) were detected either with the plant-derived chimeric PfAMA1-specific mAb4G2 (left panel) or the murine Pfs25-specific mAb4B7 (MRA28, MR4, ATCC) (right panel). d Immunoblot analysis under non-reducing conditions. Left panel: Purified VAMAX 1 (V1, 1.3 µg) was detected using a PfCSP_TSR domain-specific murine monoclonal antibody mAb6.75; right panel: Purified VAMAX 2 (V2, 1.3 µg) was detected using the PfMSP1_19-specific murine monoclonal antibody mAb5.2 (MRA94, MR4, ATCC). e LDS-PAGE (left panel, 3.4 µg) and immunoblot analysis (right panel, 1.3 µg) of an equimolar mixture of VAMAX 1, VAMAX 2 and VAMAX 6 (VM). Immunoblot analysis was performed under non reducing conditions and the equimolar VAMAX mixture was detected with the plant-derived chimeric PfAMA1-specific mAb4G2. Primary antibodies were detected with alkaline phosphatase-labeled goat anti-human or anti-mouse antisera. M: PageRuler™ Pre-stained protein ladder (Fermentas)

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