Enhanced protection of modified human acidic fibroblast growth factor with polyethylene glycol against ischemia/reperfusion-induced retinal damage in rats.
Molecular modification with polyethylene glycol (PEGylation) is an effective approach to improve protein biostability and decrease protein immunogenic activity. To create a PEGylated recombinant human acid fibroblast growth factor (rhaFGF) and improve its bio-stability, we have produced a rhaFGF mutant (rhaFGF(ser98,132)) by replacing the 98th and the 132nd cysteine residues with serine residues. The rhaFGF(ser98,132) that retains the bioactivity of rhaFGF was then site-specifically conjugated with PEG-maleimide at the 31st cysteine residue. PEGylated rhaFGF(ser98,132) has less effect than the native rhaFGF(ser98,132) on stimulating 3T3 cell proliferation in vitro; however, its biostability at a prolonged incubation under various temperatures and resistance to trypsinization were significantly enhanced, and half-life time in vivo was elongated while its immunogenicity was significantly decreased. The physiological function of PEGylated rhaFGF(ser98,132) was evaluated in a rat model of retinal ischemia/reperfusion injury, showing that in vivo supplementation of PEGylated rhaFGF(ser98,132) provided a significantly better protection than the native rhaFGF(ser98,132) against ischemia/reperfusion-induced retinal morphological changes and lipid peroxidation. The protection is probably mediated by antioxidant protective mechanisms.
Huang
Zhifeng
Z
Chinese-American Research Institute for Diabetic Complications, School of Pharmaceutical Science, Wenzhou Medical College, China.
Zheng
Qing
Q
Wu
Xiaoping
X
Su
Zhijian
Z
Xu
Hua
H
Tan
Yi
Y
Feng
Wenke
W
Li
Xiaokun
X
Cai
Lu
L
D000818:Animals; D041702:BALB 3T3 Cells / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004195:Disease Models, Animal; D004355:Drug Stability; D005260:Female; D005346:Fibroblast Growth Factors / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007429:Intraocular Pressure / Q000187:drug effects; D015227:Lipid Peroxidation / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D011092:Polyethylene Glycols / Q000378:metabolism / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D015427:Reperfusion Injury / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D012160:Retina / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D012162:Retinal Degeneration / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*
D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D005346:Fibroblast Growth Factors