Abstract:

Plasmid is a kind of popular nonviral vector in gene therapy. To produce large quantities of plasmid, an efficient large-scale purification process needs to be developed. In general, liquid chromatography is the most important tool for the analytical and preparative separation of plasmid DNA. In this study, biporous polymeric microspheres coupled with mercaptopyridine were synthesized for the purification of plasmid DNA. From the results of experiments, it can be inferred that the mechanism for the purification of the plasmid DNA with MP-BiPR is based on the hydrophobic interaction. When the load was 10 mL with the content of plasmid of 0.30 mg/mL and the content of RNA of 0.44 mg/mL, a flow rate was set up to 4 mL/min (20 cm/min). The separation was finished in 10 min, and the plasmid was completely recovered with undetectable impurities of nucleic acids and proteins. The load of the column for RNA was up to 2.2 mg/mL.

Key words: biporous polymeric microspheres stationary phase, plasmid, rapid purification , mercaptopyridine