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. Author manuscript; available in PMC: 2019 Feb 8.
Published in final edited form as: Mol Biol Rep. 2018 Jul 21;45(5):1551–1556. doi: 10.1007/s11033-018-4251-3

Figure-3. Synthetic FhSAP2 retains its main structural and antigenic properties.

Figure-3.

(A) 12.5% SDS-PAGE analysis of S-FhSAP2 was performed under reduction conditions. Lane-1 represents untreated S-FhSAP2 and lane-2 represents S-FhSAP2 treated with 2-mercaptoethanol. (B) Circular dichroism (CD) spectra of S-FhSAP2 were performed by measurements in the far-UV region (180–250 nm). A Jasco J-1500 CD spectrometer was used with protein concentration of 0.1 mg mL−1 in 50mM phosphate buffer, pH 8.0 in the 190–250 nm ranges at 25°C using 0.1-cm path-length cell. (C) ELISA was used to test the ability of S-FhSAP2 to react with specific anti-FhSAP2, anti-ESP sera or a pool of sera from animals with 4 weeks of F. hepatica infection. (D) Protein yield and expression protocol duration when FhSAP2 is expressed in E. coli vs. a cell-free expression system. *Includes a primary overnight culture of E. coli that is further used to inoculate 1 liter of culture until exponential phase, followed by 4 hours of expression induction with 0.02% L-arabinose, and purification of protein from inclusion bodies. **Estimated after purification by BCA method.