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Circumsporozoite protein (CSP), the most abundant surface protein of SPZ is the leading candidate for passive (and subunit) immunization approaches against malaria. Comprehensive assessment of the parasite-inhibitory capacity of anti-CSP monoclonal antibodies (mAbs) is an important step in advancing CSP-based immunization strategies. In this study, we employed a quantitative imaging-based motility assay to quantify the effect of anti-CSP mAbs on SPZ motility, both in vitro and in human skin.<\/jats:p><jats:p>Our assay provided a quantitative measure of mAb parasite-inhibitory capacity through measurement of the half-maximal motility inhibitory concentration (IC<jats:sub>50M<\/jats:sub>) value for anti-CSP mAbs (IC<jats:sub>50M<\/jats:sub> 2A10: 24\u00a0nM, IC<jats:sub>50M<\/jats:sub> 3SP2: 71\u00a0nM). We found a sevenfold discrepancy between the IC<jats:sub>50M<\/jats:sub> and the binding saturation concentration measured by ELISA, possibly related to the observed shedding of CSP-mAb complexes during SPZ movement. In a subset of SPZ (5%), in vitro motility was unaffected by the presence of 2A10 while 3SP2 was able to completely block movement. In our ex vivo skin explant model, SPZ proved less susceptible to anti-CSP mAbs compared to SPZ in an in vitro environment. By quantitatively assessing motility, we created a valuable tool that can be used for comprehensive assessment of anti-CSP mAb potency. Insight that will help deepen our understanding of anti-CSP mAb potency and guide selection of the most promising anti-CSP mAbs for downstream clinical development.<\/jats:p>","DOI":"10.1038\/s41598-022-22154-8","type":"journal-article","created":{"date-parts":[[2022,10,13]],"date-time":"2022-10-13T15:06:09Z","timestamp":1665673569000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Sporozoite motility as a quantitative readout for anti-CSP antibody inhibition"],"prefix":"10.1038","volume":"12","author":[{"given":"C. M.","family":"de Korne","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R.","family":"van Schuijlenburg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. C.","family":"Sijtsma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"H. 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