Fig 2: Comparison of TALON resin and magnetic Dynabeads for MVP-H6 purification. Samples were analyzed by (A) SDS-PAGE and TGX visualization, and (B) Western blot developed with a monoclonal antibody against His-tag. Legend: initial sample (In), flowthrough (FT), first wash (W1), second wash (W2), elution with 250 mM imidazol (E1) and with 2 M imidazol (E2). R and MP correspond, respectively, to samples of resin and magnetic particles after elution steps, in order to evaluate amounts of protein not eluted and still attached to the purification support. MW: molecular weight standard.

Fig 3: Solubility of MVP-H6 produced in cultures of E. coli cells induced with 1 mM IPTG, either for 3 h at 37 °C (A) or o/n at 20 °C (B). After obtaining soluble (Sol) and insoluble (Ins) cell fractions, they were submitted to SDS-PAGE and further Western blot, developed with an anti-His monoclonal antibody.

Fig 4: Solubility of MVP-H6 produced at different IPTG concentrations. Three independent cultures (A–C) were analyzed for each condition. After obtaining insoluble (Ins) and soluble (Sol) cell fractions, they were submitted to SDS-PAGE and further Western blot, developed with an anti-His monoclonal antibody.

Fig 5: Optimization of conditions for MVP-H6 purification. Purification of MVP-H6 protein with TALON Superflow resin, using different concentrations of imidazol as elution agent. Samples were analyzed by (A) SDS-PAGE and TGX visualization, and (B) Western blot developed with a monoclonal antibody against His-tag. Legend: initial sample (In), and flowthrough (FT). Numbers 20 to 500 indicate imidazol concentration (mM) used in each step. MW: molecular weight standard.