Production and Evaluation of an Avian IgY Immunotoxin against CD133+ for Treatment of Carcinogenic Stem Cells in Malignant Glioma: IgY Immunotoxin for the Treatment of Glioblastoma
Figure 2
Recombinant expression and purification of abrin A-chain. (a) Agarose gel shows the pET-28a (+) plasmid digestion containing the abrin A chain insert. Lane 1 shows the plasmid digested by restriction enzymes HindIII and XhoI. Lane 2 shows the plasmid with the abrin A chain sequence insert. (b) 12% SDS-polyacrylamide gel shows the expression of recombinant abrin A chain by E. coli BL21DE3pLysS; fractions of cultures before and after induction with IPTG are shown (lanes 1 and 2, respectively). (c) A 12% SDS-PAGE shows the products of purification of abrin with IPTG: lane 1, proteins of total bacterial lysates; lane 2, proteins not adhered to the affinity column; lanes 3 and 4, products eluted after washes; lane 5, proteins eluted in the first fraction; lane 6, proteins eluted in the second fraction; lane 7, proteins eluted in the third fraction. (d) WB of abrin: lane 1 proteins of total bacterial lysates; lane 2 proteins not adhered to the affinity column; lanes 3 and 4, products eluted after washed; lanes 5 and 6, protein eluted from the Ni-column.