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. 2021 Mar 19;12:1750. doi: 10.1038/s41467-021-21955-1

Fig. 1. Human recombinant mAbs were generated from Pfs230D1-specific single memory B cells of Malian adults vaccinated with the Pfs230D1-EPA/Alhydrogel® TBV.

Fig. 1

a VH and VL genes corresponding to each mAb. LMIV230-01 and LMIV230-02 sequences originate from the IGHV1-69 heavy chain gene but utilize different kappa chain genes. Complete V gene usage determined in Pfs230-specific memory B cells is described in Supplementary Fig. 3e, f. b Epitope binning of human anti-Pfs230D1 scFvs. The primary binding scFv is listed on the left and the competing scFv is listed on the top. Reported scores are a percentage of total binding of that antibody in the absence of a competitor scFv. Values greater that 50% display low amounts of competition, while values lower than 50% exhibit greater competition. Any experiment with >100% binding was given a score of 100, while negative values were given a score of 0. Potential epitope bins are grouped and labeled above the table. c Functional activity of each mAb, assessed by Standard Membrane Feeding Assay (SMFA) and measured as the % reduction (versus control mAb) in the number of P. falciparum NF54 oocysts in midguts of Anopheles mosquitoes (“TRA”). d LMIV230-01 and LMIV230-02 mAbs bound similarly to Pfs230D1 and (e) show high affinity to recombinant Pfs230D1 (Supplementary Fig. 5, Supplementary Table 2). f LMIV230-01 reduces P. falciparum NF54 oocyst numbers by 91.7% at 1000 µg/mL, 86.7% at 500 µg/mL, 87.7% at 250 µg/mL, and 80.3% at 60 µg/mL, while LMIV230-02 displays only modest activity with 58.7% reduction at the maximum concentration of 1000 µg/mL, in SMFA. At least three biological replicates were performed for each concentration of mAb, in the presence of intact serum containing human complement. N ≥ 20 mosquitos per assay. Average oocyst numbers in the control mosquitoes (fed with mouse IgG1 mAb targeting P. yoelii P140 protein) for each experiment were: exp. 1 = 29.73; exp.2 = 7.18; exp. 3 = 57.86; exp. 4 = 36.41; exp. 5 = 51.71, exp. 6 = 4.55; exp. 7 = 62.35; exp. 8 = 20.50, exp.9 = 8.71, exp 10 = 18.05, exp. 11 = 5.86. Negative oocyst reduction values were set to zero. Human isotype IgG1 and US human serum pool were used as additional negative controls (Supplementary Fig. 6b). Values are shown as mean ± s.e.m. g LMIV230-01 and LMIV230-02 bind to non-reduced (NR) protein extract of P. falciparum NF54 gametes purified on Nycodenz after 2 h in exflagellation medium. h LMIV230-01 binds to gametes at 7.5 µg/mL while LMIV230-02 does not bind at 7.5 µg/mL. or 30 µg/mL. Both mAbs were labeled with Alexa Fluor 488. Scale bars: 5 µM. This experiment was performed in triplicate. Source data are provided as a Source Data file.