title: High-level expression and purification of a nonmitogenic form of human acidic fibroblast growth factor in Escherichia coli.
abstract: To decrease the potential side effects of acidic fibroblast growth factor (aFGF) caused by its broad-spectrum mitogenic activity, a nonmitogenic form of aFGF (nhaFGF), which retained the cardio- and neuroprotective characters of the wild-type aFGF, was overexpressed in Escherichia coli. The expression level of nhaFGF was up to 25% of the total cellular protein. The expressed nhaFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. The mitogenic activity of the purified nhaFGF was decreased dramatically comparable to that of the wild-type aFGF (haFGF) detected by methylthiazoletetrazolium method. The purified recombinant nhaFGF was sufficiently prepared and sufficient for the following pharmacological study.
authors.affiliation: Biopharmaceutical Research and Development Center, Pharmacy College, Jinan University, Guangzhou, PR China. xp_wu@hotmail.com
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Wu
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Su
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Li
authors.forename: Qing
authors.initials: Q
authors.lastname: Zheng
authors.forename: Yadong
authors.initials: Y
authors.lastname: Huang
authors.forename: Hui
authors.initials: H
authors.lastname: Yuan
mesh_terms: D000818:Animals; D041702:BALB 3T3 Cells; D049109:Cell Proliferation / Q000187:drug effects; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D016220:Fibroblast Growth Factor 1 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D014169:Transformation, Bacterial / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D016220:Fibroblast Growth Factor 1