Blood was collected from mice via either tail vein or retro-orbital bleeds and left to clot overnight at 4 °C, and the following day sera was collected by spinning the blood at 2000 g for 15 minutes at 4 °C. Next, Maxisorp Nunc-Nucleon 96 flat bottom plates were coated with 1 μg/ml streptavidin overnight at 4 °C in dark. The following day, plates were washed in wash buffer consisting 0.05% tween20 in PBS, and were incubated for 1 hour with different biotinylated peptides listed in pertaining to diverse sections of CSP protein. The peptide sequences were as follows, CSPNterm: Biotin-Ahx-QEYQCYGSSSNTRVLNELNYDNAGTNLYNELEMNYYGKQENWYSLKKNSRSLGENDDGNNEDNEKLRKPKHKKLKQPADG; CSPRepeat: Biotin-Ahx-NANPNANPNANPNANPNANPNANP NANPNANPNANP, and CSPCterm: Biotin-Ahx-NKNNQGNGQGHNMPNDPNRNVDENANANSAVKNNNNEEPSDKHIKEYLNKIQNSSTEWSPCSVTCGNGIQVRI KPGSANKPKDELDYANDIEKKICKMEKCS. Additional peptides also carrying a Biotin-Ahx linker are described in Figure S6. Wells were blocked for 1 hour before incubating with initial sera (dilution of 1/100 times, followed by 1 in 4 serial dilutions down the plate) for one hour. After washing, anti IgG detection antibody conjugated to HRP was diluted 1/2000 times and was incubated for one hour. The plates were washed, then developed with Peroxidase Substrate Kit for 15 minutes, and the reaction was stopped using 50 μL/ well stop solution, consisting of 10% SDS in PBS. Absorbance at 405 nm (A405) was measured using an Infinite PRO Tecan plate reader, and the data was expressed as area under the curve (AUC) calculated in Prism 7 from the log(dilution) on the x axis and the A405 on the y axis, fitting a sigmoidal curve.

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