Synthesis and biological activity of polyethylene glycol-mouse nerve growth factor conjugate.
Conjugation of poly(ethylene glycol) derivatives with therapeutic proteins is a promising approach for enhancing protein stability and, therefore, effectiveness. An N-hydroxysuccinimidyl ester of fluorescein-PEG 2000 was used for chemical modification of mouse nerve growth factor (mNGF), a dimeric protein with therapeutic potential for Alzheimer's disease. The mNGF-PEG2000-fluorescein conjugate was characterized by RP-HPLC, spectrofluorometry, and SDS-PAGE and was biologically active, as determined by two independent NGF-specific assays (enhancement of ChAT activity in fetal neurons and neurite outgrowth in PC12 cells). The conjugate was not detectable by a standard NGF ELISA, suggesting a fortuitous reduction in protein recognition by antibodies.
Belcheva
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N
School of Chemical Engineering, Cornell University, 120 Olin Hall, Ithaca, New York 14853, USA.
Woodrow-Mumford
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K
Mahoney
M J
MJ
Saltzman
W M
WM
D000544:Alzheimer Disease / Q000188:drug therapy; D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D019281:Dimerization; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D004622:Embryo, Mammalian; D019793:Fluorescein / Q000737:chemistry; D051379:Mice; D020932:Nerve Growth Factor / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology; D016501:Neurites / Q000648:ultrastructure; D009474:Neurons / Q000201:enzymology; D016716:PC12 Cells; D011092:Polyethylene Glycols / Q000008:administration & dosage / Q000737:chemistry*; D051381:Rats; D013050:Spectrometry, Fluorescence; D013388:Succinimides / Q000737:chemistry
D013388:Succinimides; D011092:Polyethylene Glycols; D020932:Nerve Growth Factor; D015500:Chloramphenicol O-Acetyltransferase; C001426:N-hydroxysuccinimide; D019793:Fluorescein