A PEGylation technology of L-asparaginase with monomethoxy polyethylene glycol-propionaldehyde.
Polyethylene glycol (PEG) conjugation technology has been successfully applied to improve the performance of protein drugs. In this study, L-asparaginase was N-terminal site-specifically modified by alkylating PEG with monomethoxy polyethylene glycol-propionaldehyde (mPEG-ALD20000). The optimum reaction parameters were determined as pH 5.0, a molar ratio of mPEG-ALD2000 to L-asparaginase of 10:1, a reaction time of 16 h and temperature of 25 degrees C. PEG-L-asparaginase (PEG-L-ASNase) was isolated and purified with consecutive anion-exchange (XK, 16 x 20 cm, Q Sepharose FF) and gel-filtration (Tricorn, 10 x 600 cm, Sephacryl S-300 HR) chromatography, respectively. PEG-L-ASNase retained 43.5% of its activity and the N-terminal amino groups were modified to an extent of 3.67%.
College of Bioengineering, Chongqing University, Chongqing City, 400030, China. wangbc2000@126.com
Bochu
B
Wang
Yang
Y
Cao
Shaoping
S
Chi
Deshuai
D
Lou
D000447:Aldehydes / Q000737:chemistry*; D001215:Asparaginase / Q000737:chemistry*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D011092:Polyethylene Glycols / Q000737:chemistry*; D013696:Temperature
D000447:Aldehydes; D011092:Polyethylene Glycols; C005556:propionaldehyde; D001215:Asparaginase