Outline of the Enhanced Chemiluminescent-Slot Blot (ECL-SB) experimental procedure.

<p>Outline of the Enhanced Chemiluminescent-Slot Blot (ECL-SB) experimental procedure.</p

Range of detection of r<i>Pf</i>CSP in the ECL-SB.

<p>A: Two-fold dilutions of r<i>Pf</i> CSP were transferred onto a nitrocellulose membrane using a slot blot apparatus and the membrane was probed with the mAb 2A10. The amount of protein was determined by measuring the intensity of a specific band using the ImageJ software and was represented as integrated optical density (IOD). B: A standard curve based on the IOD values was derived using the Michaelis–Menten hyperbolic function. The linear range of antigen–antibody reaction shown in inset is obtained with 2.5–20 pg of r<i>Pf</i> CSP.</p

Relationship between ECL-Western blot (ECL-WB) and ECL-SB.

<p>The IOD values obtained from ECL-WB (A) and ECL-SB (B) analysis of two-fold dilutions of r<i>Pf</i> CSP (20 pg -1.25 pg) were overlaid (C).</p

Range of detection of <i>Pf</i> Oocyst by ECL-SB.

<p>A: Two-fold dilutions of <i>Pf</i> Oocysts were transferred onto a nitrocellulose membrane using a slot blot apparatus and probed with the mAb 2A10. The amount of protein was determined by measuring the intensity of a specific band using ImageJ software and was represented as integrated optical density (IOD). B: A standard curve based on the IOD values was derived using the Michaelis–Menten hyperbolic function. The linear range of antigen–antibody reaction shown in the inset is obtained within 1–4 <i>Pf</i> Oocysts.</p

The dynamics of circumsporozoite protein (CSP) expression on developing <i>P. falciparum</i> oocysts in the mosquito midgut.

<p>A: Midguts from 10 infected or uninfected mosquitoes were dissected on days 7–9 post feeding. Lysates were prepared by boiling the midguts in 1X SDS-PAGE gel loading dye for 5 minutes (2 midguts/10 µl dye). 10 µl of the sample was subjected to electrophoresis on 4–12% NuPAGE gel and transferred onto a PVDF membrane and probed with the mAb 2A10. No midgut was collected on the evening of day 9 due to saturation of <i>Pf</i> CSP expression levels. Lanes 1–5: <i>Pf</i> Oocysts from infected mosquitoes; Lane 6: Lysate from normal uninfected midgut (NUM). AM – Midgut samples collected around 10.00 AM; PM – Midgut samples collected around 3.00 PM. B: The amount of protein was determined by measuring the intensity of a specific band using the ImageJ software and was represented as integrated optical density (IOD).</p

r<i>Pf</i> CSP detection - Intra-assay and inter-assay variability.

<p>5pg of r<i>Pf</i> CSP in 20 µl were loaded in eight wells using a slot blot apparatus on to nitrocellulose membrane and probed with 2A10 mAb. The r<i>Pf</i> CSP and 2A10 mAb reactivity were determined by measuring the intensity of protein band using ImageJ software and were represented as IODs. An experiment with identical design was repeated on three different days and the IOD values were used to calculate intra-assay and inter-assay variability.</p

L'Auto-vélo : automobilisme, cyclisme, athlétisme, yachting, aérostation, escrime, hippisme / dir. Henri Desgranges

14 septembre 19181918/09/14 (A19,N6445)

r<i>Pf</i> CSP detection - Intra-Assay variability.

<p>* Integrated optical density was measured for 5 pg of r<i>Pf CSP</i>.</p><p>** CV (%)  =  (SD of Sample IODs/Mean) x 100.</p><p>r<i>Pf</i> CSP detection - Intra-Assay variability.</p

r<i>Pf CSP</i> detection - Inter assay Variability.

<p>* Integrated optical density was measured for 5 pg of r<i>Pf CSP</i>.</p><p>** CV (%)  =  (SD of Means/Grand Mean) x 100.</p><p>r<i>Pf CSP</i> detection - Inter assay Variability.</p

Phase 1 Study in Malaria Naïve Adults of BSAM2/Alhydrogel®+CPG 7909, a Blood Stage Vaccine against <em>P. falciparum</em> Malaria

<div><p>A Phase 1 dose escalating study was conducted in malaria naïve adults to assess the safety, reactogenicity, and immunogenicity of the blood stage malaria vaccine BSAM2/Alhydrogel®+ CPG 7909. BSAM2 is a combination of the FVO and 3D7 alleles of recombinant AMA1 and MSP1₄₂, with equal amounts by weight of each of the four proteins mixed, bound to Alhydrogel®, and administered with the adjuvant CPG 7909. Thirty (30) volunteers were enrolled in two dose groups, with 15 volunteers receiving up to three doses of 40 µg total protein at Days 0, 56, and 180, and 15 volunteers receiving up to three doses of 160 µg protein on the same schedule. Most related adverse events were mild or moderate, but 4 volunteers experienced severe systemic reactions and two were withdrawn from vaccinations due to adverse events. Geometric mean antibody levels after two vaccinations with the high dose formulation were 136 µg/ml for AMA1 and 78 µg/ml for MSP1₄₂. Antibody responses were not significantly different in the high dose versus low dose groups and did not further increase after third vaccination. <em>In vitro</em> growth inhibition was demonstrated and was closely correlated with anti-AMA1 antibody responses. A Phase 1b trial in malaria-exposed adults is being conducted.</p> <h3>Trial Registration</h3><p>Clinicaltrials.gov <a href="http://clinicaltrials.gov/ct2/show/NCT00889616">NCT00889616</a></p> </div