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2008122112ChinJBiologicalsDecember2008Vol.21No.12Q782R392-33A1004-5503200812-1080-03【】1【】1。180%。37℃25℃4℃-20℃37℃、25℃4℃-20℃。14℃-20℃。【】1StabilityofRecombinantPhage-displayedDiabetesType1VaccineSeedsWANGHai-xuan,DUANJin-mei,HUNing-zhu,etalInstituteofMedicalBiology,ChineseAcademyofMedicalSciencesandPekingUnionMedicalCollege,Kunming650118,China【Abstract】ObjectiveToobservethestabilityofrecombinantphage-displayeddiabetestype1vaccineseedsatdifferenttemperatures.MethodsStoretherecombinantphage-displayeddiabetestype1vaccineseedscontainingglycerolasstabilizeratdif-ferenttemperatures,usingthosecontainingnoglycerolascontrol.Takesamplesondifferentdaysafterstoragefordeterminationofphagetiterbyplaqueassay.ResultsThetitersofvaccineseedsdecreasedrapidlyat37and25℃,whiledecreasedslowlyat4and-20℃.Glycerolshowednosignificantlyprotectiveeffectonthevaccineseedsstoredat37,25and4℃,whileshowedsignificantlyprotectiveeffectat-20℃.ConclusionRcombinantphage-displayeddiabetestype1vaccineseedsmaybestoredforashorttimeat4℃,whileforalongtimeat-20℃,andglycerolmaybeusedasastabilizer.【Keywords】Phage-display;Diabetestype1vaccine;Seeds;Stability1。T7。T710A34410B397。10B。T72。1。1、。1.1.11152.0×1011pfu/mlE.coliBLT5403Novagen。1.2、OXOIDLTD。1.32180%Eppendorf0.5ml1。37、25、4-20℃。1.411.4.1E.coliBLT5403LBAmpr37℃5mlLB37℃150r/min50μl5mlLBAmpr37℃150r/min3hA600=0.6~0.8。863“I”2006AA02Z4C0“1”2006SG05.650118.E-mail:huyunz@21cn.com1080··2008122112ChinJBiologicalsDecember2008Vol.21No.121.4.2LB1050℃。3108、109、1010。1.4.33~5。A600=0.6~0.8E.coliBLT5403250μl,100μl,37℃LB37℃3h。。pfu/ml=××10×2。2.2.137℃37℃71011pfu/ml109pfu/ml7108pfu/ml1。137℃1Tab1.Titersofphage-displayeddiabetes1vaccineseedsstoredat37℃forvariousdaysTiters×108pfu/mlonvariousdaysafterstorage234567Glycerol-free1199658233420Glycerol-containing4362341Adjustedvalue*86124682*Adjustedvalue=Titerofglycerol-containingvaccine×2.2.225℃25℃15109pfu/ml108pfu/ml2。225℃1Tab2.Titersofphage-displayeddiabetes1vaccineseedsstoredat25℃forvariousdaysTiters×108pfu/mlonvariousdaysafterstorage3579111315Glycerol-free8904004216542335Glycerol-containing1050500113662Adjustedvalue*2100100022612124*Adjustedvalue=Titerofglycerol-containingvaccine×2.2.34℃4℃471010pfu/ml37℃25℃3。34℃1Tab3.Titersofphage-displayeddiabetes1vaccineseedsstoredat4℃forvariousdaysTiters×108pfu/mlonvariousdaysafterstorage281622324647Glycerol-free1480600400400400400300Glycerol-containing10009009001001009073Adjustedvalue*200018001800200200180146*Adjustedvalue=Titerofglycerol-containingvaccine×2.2.4-20℃-20℃601010pfu/ml90107pfu/ml901010pfu/ml150108pfu/ml4。4-20℃1Tab4.Titersofphage-displayeddiabetes1vaccineseedsstoredat-20℃forvariousdaysTiters×108pfu/mlonvariousdaysafterstorage306090120150Glycerol-free5001000.600.140.12Glycerol-containing100020022092Adjustedvalue*2000400440184*Adjustedvalue=Titerofglycerol-containingvaccine×2.3.-TTVaccineVaccineVaccineVaccine1081··2008122112ChinJBiologicalsDecember2008Vol.21No.1210778aa29、36、49、71、87、123、242、278aa36、278VR2、VR3、VR4、VR5、VR6VR9。VP7。G10VP7。VP7Aaa87-101、Baa141-150、C208-2213VR5、VR7、VR8ACVP785%910。LLRG10RV96.7%RV40.0%~66.7%VP7。3LLR14G10RVAaa8797.5%G1042.5%~62.5%G1、G2、G3、G452.5%、52.5%、52.5%55.0%。G10LLRG1~44G10G1、G2、G3、G4VP7。LLRVP7LLRVP7LLR。1ParasharUD,GibsonCJ,BresseJS,etal.Rotavirusandseverechildhooddiarrhea.EmergInfectDis,2006,122:304-306.2MohanKV,KulkarniS,GlassRI,etal.AhumanvaccinestrainoflambrotavirusChineseNSP4gene:completenucleotidese-quenceandphylogeneticanalyses.VirusGenes,2003,262:185-192.3Higo-MoriguchiK,AkahoriY,IbaY,etal.Isolationofhumanmonoclonalantibodiesthatneutralizehumanrotavirus.JVirol,2004,787:3325-3332.4..1.20041351-1362.5MatthijnssensJ,RahmanM,YangX,etal.G8rotavirusstrainiso-latedintheDemocraticRepublicofCongobelongtotheDS-1-likegenogroup.JClinMicrobiol,20065:1801-1809.6RaoCD,GowdaK,ReddyBS.SequenceanalysisofVP4andVP7genesofnontypeablestrainsidentifiesanewpairofouterproteinsrepresentingnovelPandGgenotypesinbovinerotaviruses.Virolo-gy,2000,2761:104-113.7.AVP7.2003172165-168.8GreenKY,HoshinoY,IkegamiN.Sequenceanalysisofthegeneencodingtheserotype-specificglycoproteinVP7oftownewhu-manrotavirussetypes.Virology,1989,1682:429-433.9DasM,DunnSJ,WoodGN,etal.BothsurfaceproteinsVP4andVP7ofanasymptomaticneonatalrotavirusstrain1321havehighlevelsofsequenceidentitywiththehomologousproteinofaserotype10bovingrotavirus.Virology,1993,1941:374-379.10GreenKY,SearsJF,TaniguchiK,etal.Predictionofhumanro-tavirussetypebynucleotidesequenceanalysisoftheVP7proteingene.JVirol,1988,625:1819-1823.2008-04-286。、。37℃、25℃1。78。4℃、-20℃-20℃9。14℃-20℃。1..2005184350-353.2..200829160-63.3.cDNA.2007255406-410.4..2008484498-502.5..2007342128-129.6..,2006233193-196.7..2007241043-44.8.H5N1.20082418-13.9..2006285413-417.2008-07-111082··
本文标题:噬菌体展示的1型糖尿病疫苗种子的稳定性
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