ELISAs against Pfs25 recombinant protein were performed using standardized methodology as previously described for murine (10 (link), 73 (link)) and human (69 (link), 74 (link)) samples, except that plates were blocked with StartingBlock™ T20 (PBS) Blocking Buffer (ThermoFisher Scientific,UK).
For Pfs28 endpoint ELISA, Nunc-Immuno maxisorp plates were coated with recombinant Pfs28 protein, produced from the Drosophila S2 cells. Indicated serum samples were added in duplicates and diluted 3 fold down the plate, followed by the same procedure as for the Pfs25 standardized ELISA. Optical density (OD) was read at 405 nm using an ELx800 absorbance microplate reader (Biotek, UK). The endpoint titer is defined as the X-axis intercept of the dilution curve at an absorbance value ( ± three standard deviations) greater than the OD for a negative control serum sample.
For Pfs28 endpoint ELISA, Nunc-Immuno maxisorp plates were coated with recombinant Pfs28 protein, produced from the Drosophila S2 cells. Indicated serum samples were added in duplicates and diluted 3 fold down the plate, followed by the same procedure as for the Pfs25 standardized ELISA. Optical density (OD) was read at 405 nm using an ELx800 absorbance microplate reader (Biotek, UK). The endpoint titer is defined as the X-axis intercept of the dilution curve at an absorbance value ( ± three standard deviations) greater than the OD for a negative control serum sample.
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