Figure 2
From: Mimotopes selected with neutralizing antibodies against multiple subtypes of influenza A
The purification of recombinant multi-mimotope of influenza A virus. a: The multi-mimotope was expressed in soluble form in bacteria and purified with affinity chromatography (lane 1: protein marker: 116.0, 66.2, 45.0, 35.0, 25.0, 18.4, 14.4KDa; lane 2: multi-mimotope gene was transferred to bacteria and induced with IPTG; lane 3: supernatant of ultrasound-broken bacteria; lane 4: pellete of ultrasound-broken bacteria; lane 5: flow-through of supernatant loaded on Ni2+-NTA-resin; lane 6: first 0.25 ml elution from 0.25 ml Ni2+-NTA-resin with 60 mM imidazole; lane 7: second 0.25 ml elution from Ni2+-NTA-resin with 60 mM imidazole; lane 8-9: elution from Ni2+-NTA-resin with 1 M imidazole). b: The optimized concentration gradient between 5 and 60 mM imidazole for affinity chromatography (lane 1: protein marker: 97.4, 66.4, 43.0KDa; lane 2-3: elution from Ni2+-NTA-resin with 5 mM imidazole; lane 4-5: elution from Ni2+-NTA-resin with 10 mM imidazole; lane 6-7: elution from Ni2+-NTA-resin with 30 mM imidazole; lane 8-9: elution from Ni2+-NTA-resin with 60 mM imidazole). c: The multi-mimotope was purified with repeat affinity chromatography (lane 1: protein marker: 116.0, 66.2, 45.0, 35.0, 25.0, 18.4, 14.4KDa; lane 2: supernatant of ultrasound-broken bacteria; lane 3: multi-mimotope purified by repeat affinity chromatography).