Expression of recombinant human nerve growth factor in Escherichia coli.
Nerve growth factor (NGF) is a neurotrophic factor for basal forebrain cholinergic neurons and may be of benefit in neurodegenerative diseases of humans. A method is described to obtain significant amounts of biologically active recombinant human NGF (rhNGF) in one step. RhNGF was expressed in E. coli and the majority of the protein accumulated in inclusion bodies. It was immunoprecipitated by a serum against mouse NGF. Solubilization of the inclusion bodies was done in 3M guanidine HCl and renaturation was effected by dilution and air oxidation in the presence of 6 microM CuSO4. Recoveries were 10-12 micrograms of rhNGF per ml of bacterial suspension. Its biological activity was tested in a bioassay system employing sympathetic chick embryo ganglia and was inhibited by the monoclonal antibody 27/21 against mouse NGF.
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INSERM U 298, Centre Hospitalier Régional et Universitaire, Angers, France.
D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D001681:Biological Assay; D002642:Chick Embryo; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005728:Ganglia, Sympathetic / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000096:biosynthesis* / Q000494:pharmacology; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D015183:Restriction Mapping; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D000911:Antibodies, Monoclonal; D009414:Nerve Growth Factors; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins