1. Physicochemical Characterization of Recombinant Human Nerve Growth Factor Produced in Insect Cells with a Baculovirus Vector
- Author
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Jodi Fausnaugh, Natalie Saldou, Hardy Chan, Daryl K. Eggers, Eric Osen, Ken Straub, Jack Lile, Frank H. Collins, Joan Chow, Leo Gu, Raymond R. Townsend, Kurt Jarnagin, Hi-Shi Chaing, Binh T. Nguyen, Jim W. Barnett, and Lisa Erdos
- Subjects
Glycosylation ,Insecta ,Genetic Vectors ,Molecular Sequence Data ,Enzyme-Linked Immunosorbent Assay ,Biology ,Biochemistry ,Cell Line ,law.invention ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,law ,Native state ,Animals ,Humans ,Amino Acid Sequence ,Isoelectric Point ,Nerve Growth Factors ,Peptide sequence ,Chromatography, High Pressure Liquid ,chemistry.chemical_classification ,Isoelectric focusing ,Recombinant Proteins ,Amino acid ,Molecular Weight ,Isoelectric point ,chemistry ,Cell culture ,Recombinant DNA ,Baculoviridae ,Plasmids - 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.
- Published
- 1991
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