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. 2004 Jul;42(7):2966–2976. doi: 10.1128/JCM.42.7.2966-2976.2004

FIG. 10.

FIG. 10.

Detection of intimin and EspA from pathogenic E. coli with monoclonal scFvs. (A and B) Outer membrane fractions were prepared from pathogenic E. coli serotypes O127:H6 (α intimin, lane 1), O111:NM (β1 intimin, lane 2), O157:H7 (γ1 intimin, lane 3), O86:H34 (β2 intimin, lane 4), a nonpathogen K-12 strain (lane 5), and an O157:H7 intimin (−) mutant DM3 (lane 6) and then resolved on a 10% (wt/vol) SDS-PAGE gel (7.5 μl/lane, Coomassie blue-stained gel shown in panel A). A Western blot of the gel was developed with I4 (panel B, 5 ml of culture supernatant/blot). (C) Whole-cell preparations of E. coli O157:H7 were prepared and resolved on a 12% (wt/vol) SDS-PAGE gel (10 μl/lane). Lane 7 shows Coomassie blue staining, and lane 8 shows a Western blot developed with E1 (affinity-purified scFv from 25 ml of culture). ScFv binding was detected with anti-c myc antibody.