A Novel Structurally Stable Multiepitope Protein for Detection of HCV

Figure 2

Expression and purification of rMEHCV. (a) Cell lysates from a selected clone induced by IPTG visualized on a Coomassie Blue stained 12% SDS-PAGE. Lane 1, cell lysates from uninduced culture. Lanes 2–5, cell lysates from induced culture after 2, 4, 6, and 8 h, respectively; Lane M, unstained protein molecular weight marker (Fermentas Life Sciences). The arrow indicates the position of the band corresponding to rMEHCV. (b) Fractions collected after Ni-NTA chromatography visualized on 12% SDS-PAGE. Lane M, molecular weight markers (Weight Standard, Broad Range, BioRad). Lane 1, flow-through, Lanes 2–4, purified rMEHCV after elution with 500 mM imidazole. (c) Western blot analysis of the purified rMEHCV. The monoclonal antibody, anti-polyhistidine alkaline phosphatase conjugate, was used after dilution 1 : 1000 in PBS (pH 7.2). Lane M, PAGE™ Ruler (Fermentas Life Science). Lanes 1–4 correspond to the same fractions shown in (b).

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