A Plant-Produced Pfs25 VLP Malaria Vaccine Candidate Induces Persistent Transmission Blocking Antibodies against Plasmodium falciparum in Immunized Mice

Contains fulltext : 128628.pdf (publisher's version ) (Open Access

Pfs25-CP VLP purity and identity.

<p>(A) Deduced amino acid sequence of Pfs25-CP with Pfs25 sequence underlined. Amino acids boxed in the sequence were identified by N-terminal sequencing of the SDS-PAGE bands indicated. A Coomassie stain of an SDS-PAGE gel highlights the Pfs25-CP fusion polypeptide (arrowhead ‘a’) and CP monomer polypeptides (arrowheads ‘b’ & ‘c’). N-terminal sequencing of ‘a’ identified the first 5 amino acids of Pfs25, while sequencing of ‘b & c’ identified residue 26 of AlMV CP. (B) Western blot analysis of Pfs25-CP VLPs with an anti-Pfs25 mAb (left panel) and an anti-AlMV CP polyclonal serum (right panel).</p

Evaluation of TB activity in immunized mouse sera by SMFA: one vaccine dose.

a<p>The proportion (percentage) of mosquitoes infected.</p>b<p>Median number of oocysts per mosquito (range).</p>c<p>% reduction = (mean control oocyst – mean test oocyst) ÷ mean control oocyst)*100.</p>d<p>Control.</p>*<p>CP – coat protein.</p

Pfs25-CP VLP particle analysis.

<p>(A) Negative stain transmission electron micrograph of Pfs25-CP VLPs shows highly uniform particles of 19.3±2.4 nm in diameter. (B) Transmission electron micrograph of Pfs25-CP VLPs labeled with anti-Pfs25 and gold-labeled anti-mouse antibodies confirms the presence of Pfs25 on the particles. (C) DLS histogram showing a narrow size distribution for Pfs25-CP VLPs. The average hydrodynamic radius is ∼14 nm with a polydispersity of <15%. (D) Analytical SEC showing a single, major eluting species confirmed by Western blot analysis (not shown) to be Pfs25-CP VLP. The void volume of the SRT 1000 column (range 7.5 MDa –50 kDa) is indicated by (a) molecular weight standards indicated by (b) for thyroglobulin, (c) for BSA and (d) for uracil.</p