Figure 1

Generation of mE-ERH and isolation of EGF_ Pf MSP4-specific monoclonal hybridoma cell lines. (A) Expression cassette of the pTRAkc_mE-ERH vector used for the expression of mE-ERH. The vector contains the Cauliflower mosaic virus 35S promoter (CaMV 35S promoter), a 5′ UTR of the Chalcone synthase of Petroselium crispum (5′ UTR (Cs)), a signal peptide sequence followed by mE (the multi-EGF_MSP protein as explained in (B) and (C)), which was inserted using NcoI/NotI restriction sites, a 6-fold histidine-tag for purification, an ER-retention signal and the CaMV 35S terminator flank by scaffold attachment regions (SAR). (B) Components of the mE-ERH multidomain fusion protein. The protein consists of the EGF-like domains of MSP1-19, both of MSP8, and MSP4. (C) Peptide-sequences of the EGF-like domains of the MSPs and corresponding PlasmoDB accession numbers. (D) Reactivity of the parental polyclonal hybridoma cell line towards mE-ERH and its subdomains. The three selected monoclonal hybridoma cell lines 2.48, 2.44 and 2.7 show exclusive reactivity towards EGF_PfMSP4.