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. 2016 Mar 14;2(5):313–321. doi: 10.1021/acsinfecdis.6b00026

Figure 5.

Figure 5

Detection of HAstV-2 capsid protein by scFv PL-2. (a) Reducing SDS-PAGE analysis of purified proteins. Lanes: M, molecular weight marker; 1, mAb PL-2; 2, scFv PL-2; 3, negative control mAb NegC. (b) Anti-Strep-tag Western blot detection of scFv PL-2, which contains a Strep-tag (lane 4). (c) SDS-PAGE (left) and anti-His-tag Western blot (right) analyses of wheat germ extracts containing recombinant HAstV-2 capsid protein (C) or wheat germ extract alone (−). (d) SDS-PAGE (left) and anti-His-tag Western blot (right) analyses of immunoprecipitation experiments using scFv PL-2 and wheat germ extracts containing recombinant HAstV-2 capsid protein (C) or wheat germ extract alone (−). (e) ELISA detection of antibody binding to HAstV-2 capsid protein. Wells were coated with wheat germ extracts containing recombinant HAstV-2 capsid protein (+ Capsid) or wheat germ extract alone (− Capsid). Binding was detected by a HRP-conjugated goat anti-mouse IgG secondary antibody (for full-length mAbs) or HRP-conjugated Strep-Tactin (for scFv PL-2). Experiments with mAb PL-2 and scFv PL-2 were performed in triplicate. Due to limited amounts of wheat germ extract samples, the negative control experiments with mAb NegC or no primary antibody were performed in duplicate. Error bars represent the standard deviation.