title: Purification of Escherichia coli L-asparaginase mutants by a native polyacrylamide gel electrophoresis.
abstract: The antigenicity of L-asaparaginase (L-ASP) has been problematic for the treatment of leukemia for many years. In order to establish a relationship between the antigenic epitope of L-asparaginase and its antigenicity, several L-asparaginase mutants (mL-ASPs) are constructed and expressed. To effectively purify these enzyme mutants for further investigation, a native preparative polyacrylamide gel electrophoresis is developed. The simplicity and reproducibility of this approach permits the purification of different mutants from the crude enzyme extracts, with a sufficient activity to perform immunological and biological studies. Furthermore, the newly developed method is efficient and cost-effective compared with other methods, such as column chromatography and affinity chromatography. As a result, the enzyme mutants with specific activity of 300 approximately 400 U/mg are obtained by the single-step purification with a high degree of purity.
authors.affiliation: The School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, People's Republic of China.
authors.forename: Yujun
authors.initials: Y
authors.lastname: Wei
authors.forename: Jianhua
authors.initials: J
authors.lastname: Chen
authors.forename: Ruibo
authors.initials: R
authors.lastname: Jia
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.forename: Wutong
authors.initials: W
authors.lastname: Wu
mesh_terms: D001215:Asparaginase / Q000235:genetics / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology*; D009154:Mutation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D001215:Asparaginase