[Analysis of plasmid pUDKH by anion-exchange high performance liquid chromatogaphy].
pUDKH, a new plasmid DNA comprising human hepatocyte growth factor (HGF), was developed as a potential genetherapeutic product for peripheral arterial disease. An anion-exchange high performance liquid chromatographic (HPLC) method was developed to determine the proportion of plasmid DNA in both opened and supercoiled forms. A TSK gel DNA-NPR column was equilibrated with 50% buffer A (20 mmol/L Tris-HCl, pH 8.8) and 50% buffer B (buffer A with 1 mol/L NaCl added). pUDKH was analyzed by using a gradient HPLC with a mobile phase from 50% B to 80% B and with a UV detection at 260 nm. The flow rate was 0.5 mL/min. The method was also used to monitor the production procedures.
Liu
Jia
J
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Zhang
Qinglin
Q
Bi
Jianjin
J
Wang
Xiaona
X
Gong
Ping
P
Ha
Xiaoqin
X
Wang
Zhengping
Z
Wu
Zuze
Z
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D017228:Hepatocyte Growth Factor / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000032:analysis* / Q000235:genetics; D011995:Recombination, Genetic
D018076:DNA, Complementary; D017228:Hepatocyte Growth Factor