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Figure 2 | Veterinary Research

Figure 2

From: A hybrid protein containing MSP1a repeats and Omp7, Omp8 and Omp9 epitopes protect immunized BALB/c mice against anaplasmosis

Figure 2

Immunogenicity of the hybrid protein MSP1a/OMP7/9. A Expression of the hybrid MSP1a/OMP7/9 protein after IPTG induction. Left panel: Supernatants of the E. coli strain DH5αF’IQ growth on 15% SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Line 1: without IPTG at 0 h; line 2: after 8 h with IPTG; and line 3: 16 h after IPTG. MW—Molecular weight. B SDS-PAGE of hybrid MSP1a/OMP7/9 protein by affinity chromatography using Ni–NTA superflow 6xHis tagged (Qiagen, Brazil) (For detail, see “Materials and methods”). The purification was performed using an MCAC buffer (20 mM tris pH 7.9, 0.5% w/v NaCl and 10% v/v of glycerol), adjusted in pH 8.0 for binding, pH 6.0 for lavage and pH 3.0 for elution. C Western blotting (WB) with anti-HisG (Invitrogen) to characterize the molecular weight of the hybrid protein. D Western blotting (WB) of the hybrid protein to determine the antigenicity from anaplasmosis infected calf serum (left panel) and normal calf serum (right panel). E Immunogenicity of hybrid protein MSP1a/OMP7/9 after three immunization series emulsified with ISA adjuvant (MONTANIDE™ ISA 50 V2, Seppic, Brazil).

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