Purification of plasmids by triplex affinity interaction

摘要: Production of pharmaceutical grade plasmid DNA is an important issue in gene therapy. We developed a method for affinity purification plasmids by triple helix interaction. This based on sequence-specific binding oligonucleotide immobilized large pore chromatography support to target sequence the plasmid. Using design criteria derived from thermodynamic data, we produced 15mer which binds strongly under mildly acidic conditions. Plasmid was purified clarified Escherichia coli lysate incubation with beads at pH 5.0 and high NaCl concentration. After extensive washing beads, eluted alkaline buffer. The showed no RNA or cell contamination HPLC analysis total protein concentration reduced considerably. Due its mechanical stability porosity this can be used continuous process, has potential scale up.