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. 2023 Aug 22;14:1222267. doi: 10.3389/fimmu.2023.1222267

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Copyright © 2023 Ferguson, Delgado, McBride, Orbe, La Vake, Caimano, Mendez, Moraes, Schryvers, Moody, Radolf, Weiner and Hawley

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Immunoreactivity profiles of BamA ECL4 mAbs. (A) Titration ELISAs of the mAbs against native and irrelevant peptides (magenta and grey, respectively) and PfTrx^BamA/ECL4 and PfTrx ^Empty (blue and black, respectively). (B) mAb AUC values calculated for native peptide and PfTrx^BamA/ECL4. (C) Immunoblot reactivities against graded nanogram amounts of PfTrx ^BamA/ECL4 (21.4 kDa) or the BamA β-barrel (44.9 kDa). Based on the MW, PfTrx^BamA/ECL4 and BamA β-barrel were loaded at a 2:1 ratio, with 1 ng corresponding to 0.047 and 0.022 picomoles, respectively. PfTrx^Empty (200 ng) and nonspecific mAb (IGX6939) served as specificity controls. Immunoblots were done using mAbs at 4 μg/ml. (D) A rat polyclonal BamA ECL4 (residues 568–602) antiserum (10) (diluted 1:1,000) was used as a comparator. (E) Immunoblot reactivity of IGX7141 mAb (4 μg/ml) against graded nanogram amounts of Nichols (top) and Mexico A PfTrx^BamA/ECL4 (bottom) variants.