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. 2020 Jun 30;11:1279. doi: 10.3389/fimmu.2020.01279

Figure 2.

Figure 2

Construction and characterization of heterologous chimeric antigen NCz-SEGN24A. (A) Schematic representation of NCz-SEGN24A. Bacterial SAg mutant SEGN24A was fused to the C-terminal of the N-terminal domain of parasite cruzipain (Nt-Cz). A thioredoxin (Trx) tag was added to fused NCz-SEGN24A in its N-terminal, and histidine tag (His-tag) was added at the C-terminal. (B) Chromatogram of the NCz-SEGN24A purification with Superdex 200 molecular exclusion (SEC). (C) Analysis of recombinant NCz-SEGN24A by SDS-PAGE. The 69-kDa protein was detected using Coomassie blue staining after purification by Ni2+-NTA column followed by SEC (Lane 2). Arrow indicates NCz-SEGN24A band. (D) Immunochemical identity by Western blot. Domain-specific polyclonal antibodies were used as primary antibodies. SDS-PAGE gels were loaded as follows, lines: 1- MWM, 2- SEGN24A (left, 27 kDa) or Nt-Cz (right, 23 kDa), 3- NCz-SEGN24A (69 kDa), arrows point at NCz-SEGN24A band.