Figure 2: Expression and purification of recombinant proteins. The EGFRvIII, MAGE and GLEA recombinant proteins were produced in E. coli BL21 and Rosetta strains (a) and purified by affinity chromatography using Ni-NTA resin (b). MW: PageRuler Prestained Protein Ladder (Fermentas, Thermo Scientific); MW’: Molecular Weight Marker (MW 14,000-66,000; Sigma). a) 1, bacterial extract before EGFRvIII induction by IPTG; 2, bacterial extract after overnight induction by 0.5 mM IPTG; 3, bacterial extract before MAGE induction by IPTG; 4, bacterial extract after 4 h induction by 0.5 mM IPTG; 5, bacterial extract before GLEA induction by IPTG; 6, bacterial extract after 4 h of induction by 0.2 mM IPTG. b) 1, EGFRvIII recombinant protein purified from bacterial lysate under denaturing conditions (8 M urea); 2, MAGE recombinant protein purified from bacterial lysate under native conditions; 3, GLEA recombinant protein purified from bacterial lysate under denaturing conditions (8 M urea).