title: Physicochemical characterization of recombinant human nerve growth factor produced in insect cells with a baculovirus vector.
abstract: Recombinant human nerve growth factor (rhNGF) secreted by insect cells was purified by ion-exchange and reversed-phase chromatography to near homogeneity. The N-terminus of the secreted molecule was analogous to that of mouse salivary gland NGF. In its native conformation, the insect cell produced rhNGF molecules were homodimers consisting of 120 amino acid polypeptide chains. Mature rhNGF was found not to be significantly glycosylated (less than 0.08 mol of N-acetylglucosamine/mol of protein). The rhNGF was homogeneous with regard to molecular weight and amino acid sequence. Isoelectric focusing resolved the rhNGF into one major and one minor component. Because rhNGF from insect cells can be obtained in large quantities, purified to near homogeneity, and is similar to natural NGF with regard to physicochemical properties and biological activity, it is suitable for further evaluation in animal models as a therapeutic molecule for neurodegenerative diseases such as Alzheimer's disease.
authors.affiliation: Institute of Bio-Organic Chemistry, Syntex Research, Palo Alto, CA 94303.
authors.forename: J
authors.initials: J
authors.lastname: Barnett
authors.forename: J
authors.initials: J
authors.lastname: Chow
authors.forename: B
authors.initials: B
authors.lastname: Nguyen
authors.forename: D
authors.initials: D
authors.lastname: Eggers
authors.forename: E
authors.initials: E
authors.lastname: Osen
authors.forename: K
authors.initials: K
authors.lastname: Jarnagin
authors.forename: N
authors.initials: N
authors.lastname: Saldou
authors.forename: K
authors.initials: K
authors.lastname: Straub
authors.forename: L
authors.initials: L
authors.lastname: Gu
authors.forename: L
authors.initials: L
authors.lastname: Erdos
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000502:physiology*; D006031:Glycosylation; D006801:Humans; D007313:Insecta / Q000166:cytology* / Q000378:metabolism; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D009414:Nerve Growth Factors; D011994:Recombinant Proteins