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SODSODPEGSODSomack6PEG20000PEGFujita72000PEGPEGSODPEGPEGSODPEGSOD8PEGSOD6000PEG85.4%SODPEG-SODPEGSODSOD86000PEGSOD70℃3h65.7%SOD37.2%pH=4.4pH=113h82.6%87.1%SOD33.8%25.9%3h73.3%79.7%SOD32.1%17.4%1.J.1998204357-363.2JadotGVailleAMaldonadojetal.ClinicalPharmacokineticsandDeliveryofBovineSuperoxidedismutaseJ.ClinPharmcokinet199528117-19.3.J.1989163203-205.4.M.2.1997.217-223.5.PEGD.20013.6Somack-RSaifer-MGetal.Preparationoflong-actingsuperoxidedis-mutaseusinghighmolecularweightpolyethyleneglycol41000-72000daltonsJ.Free-Radic-Res-Commun199112-13Pt2553-562.7TakuyaFujitaMakiya.TargeteddeliveryofhumanrecombinantsuperoxidedismutasebychemicalmodificationwithMono-andpolysaccharidederivativesJ.TheJournalofPharmacologyandExperimentalTherapeuics19922633971-978.8.J.1995212139-141.150020rhG-CSFrhG-CSF1.8×107U/ml0.9×108/mgQ786A1004-311X200304-0030-022002-11-042003-02-131970-rhG-CSFrhG-CSF1-3200110-20024rhG-CSF11.1rhG-CSF63%1.2DTTMTTGSSGGSHTris1.31.3.18mol/L20mmol/LDTT20mmol/LTrisHClpH8.216000r/min20min10mg/ml1.3.21.3.2.1①20ml980ml0.3mmol/LGSH20mmol/LTrisHClpH8.24℃20h②0.15mmol/LGSSG0.3mmol/LGSH①③0.3mmol/LGSSG0.3mmol/LGSH①④0.3mmol/LGSSG0.15mmol/LGSH①⑤0.3mmol/LGSSG①1.3.2.2A1.3.2.1B1.3.2.120ml80ml2mol/L4℃1h900ml4℃20h1.3.2.31.3.2.212h18h24h30h36h1.3.2.41.3.2.11.3.2.21.3.2.30.1mg/ml0.2mg/ml0.3mg/ml0.5mg/ml1.3.3201.3.4rhG-CSF1.3.5rhG-CSF2000rhG-CSFNFS-60/MTT22.10.3mmol/LGSSGrhG-CSF1.76×107U/ml11rhG-CSFrhG-CSF×107U/ml123①0.510.560.520.53②0.730.690.710.71③1.111.161.161.14④1.491.441.421.45⑤1.791.721.761.762.22rhG-CSFrhG-CSF×107U/ml1231.791.721.761.761.751.761.711.742rhG-CSF3rhG-CSFhrhG-CSF×107U/ml123121.391.311.361.35181.711.651.661.67241.881.871.921.89301.691.721.681.70361.231.201.191.212.31343020038BIOTECHNOLOGYVol.13No.430═════════════════════════════════════════════════════════════════Aug.20034℃rhG-CSF24h1.89×107U/mlrhG-CSF32.4rhG-CSF0.3mg/ml0.5mg/ml0.1mg/ml0.2mg/ml0.2mg/ml4rhG-mg/mlCSFmg/mlrhG-CSF×107U/ml1230.10.960.980.940.960.960.21.891.871.931.900.950.32.742.682.702.710.900.53.883.863.973.900.78U/mg5×107U/ml×108U/mg11.930.9621.940.9731.950.971.940.972.5rhG-CSF1.94×107U/ml0.97×108U/mg33.1GSSG/GSH0.3mmol/LGSSG41mmol/LGSSG/1mmol/LGSHDTT20mmol/L3.224h18h20h24h30h21h22h25h26h3.30.15mg/ml-0.25mg/ml3.4rhG-CSF1.J.1995183124-127.2RudolphRZettimeisslGJaenickeRJ.Recon-stitutionoflacticdehydro-genasenonco-valentaggregationvs.reactivation.2.Re-activationofirrev-ersiblydenaturedaggre-gatesJ.Biochemistry1979185572-5575.3BuchnerJPastanIBrinkmannU.Amethodforincreasingtheyieldofproperlyfoldedrecom-biantproteinssingle-chainimmunotoxinsfromrenat-urationofbacterialinclusionbodiesJ.AnalBiochem1992205263-270.4.J.1999205221-223.150020E.colirhG-CSFDH5αpH30g/L30%Q786A1004-311X200304-0031-022002-11-042003-02-171971-RecombinantHumanGranulo-cyteColonyStimulatingFactorrhG-CSFrhG-CSFDH5α11.1rhG-CSFE.coliDH5OXOIDSigma40LNBS1.21.2.1LB1.5%50µg/ml30℃24h5mlLB50µg/ml30℃OD600nm0.4-0.642℃3hSDS-PAGE11.2.2NBS40L18L10%V/V30℃42℃DO240%4mol/L4mol/LpHpH1.2.3ALB1BLB+M91C5g/L10g/L5g/L5g/L5g/LNa2HPO412H2O18g/L5g/L1g/L0.5g/L0.5g/LZnCl24H2O2g/LCoCl26H2O2g/LFeSO416H2O4g/LH2BO35g/LMnCl24H2O1.6g/L1ml/L1.2.4721600nmOD0.2-0.7rhG-CSF15%SDS-PAGE22.1ABC20L1C1rhG-CSF%g/LA14.55.1B18.38.6C24.715.62.2pHpH6.67.07.44h42℃3h2pH7.0rhG-CSF2pHpHrhG-CSF%g/L6.618.819.17.027.321.67.426.912.62.342℃34h3hrhG-CSF%g/L229.810.1429.322.8613.930.62.442℃1hrhG-CSF42hrhG-CSF3h42℃3h1343120038BIOTECHNOLOGYVol.13No.431═════════════════════════════════════════════════════════════════Aug.2003