Production and Evaluation of a Recombinant Chimeric Vaccine against Clostridium botulinum Neurotoxin Types C and D
Figure 3
SDS-PAGE and Western blot analysis of protein expression and purification.
(A) SDS-PAGE of the whole E. coli cell extract after 3 h of induction of both constructs. Left, 1: Untransformed E. coli BL21 (DE3) Star; 2: E. coli BL21 (DE3) Star transformed with pAE/hcc-h cd before induction; 3: E. coli BL21 (DE3) Star transformed with pAE/hcc-h cd after induction. Right, 1: Untransformed E. coliBL21 (DE3) Star; 2: E. coli BL21 (DE3) Star transformed with pAE/ltb-h cc-h cd before induction; 3: E. coli BL21 (DE3) Star transformed with pAE/ltb-h cc-h cd after induction. (B) SDS-PAGE to evaluate the purity and integrity of purified proteins after Ni-affinity chromatography. Left, rH CC. Right, rH CD. All lanes designated “1” are untransformed E. coli BL21 (DE3) Star, and the lanes designated “2” are the purified antigens. (C) Anti-his Western blot to characterize the purified proteins. 1: Spectra Multicolor Broad Range Protein Ladder (Thermo Scientific), 2: Untransformed E. coli BL21 (DE3) Star, 3: rLTB/H CC/H CD, 4: rH CC/H CD.