基因工程乙肝新型疫苗-汉逊酵母表达系统-的开发

©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(),X(,100050):HBsAg,(CHO,C127)HBeAgHBsAg30adw2HBsAg;MOXFMD;,100g;,HBsAg400g/ml,HBsAg60g/ml,10,HBsAg,,:;;:R37312+1:A:100525673(2001)20320047208315(HBV)1,2000,,,,1985,90CHOMSD(YDV)1992,,5685%90%,10%1%,,9,,,,,,,,1(HBV)HBVDNA,1,HBV4(ORF),,HBcHBePolHBsXmRNAs,,1971S.Krugman122nm(HBsAg),HBVHBsAgSORF,,(S),(M)(L),CHB2sAg226,(S),GP27(27kD)P24(24kD),146(Asn)SNPreS255M,PreS24Asn,MGP36GP33;MN108109L,GP42GP39,HBsAg6HBsAg22nm,S,MLHBsAg14(Cys),742001293X:(19222),,,,,©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.:,;Fig.1StructureofHBVgenome2PreS,,,HBsAg,,80Neurath2MilichPreS,DNAPreSS,,,Michel3SPreS2CHO,S+PreS2,PreS2,20%25%Pasteur2MerieuxGen2HevacBGp2943PreS2,HBs,4160%,290%2PreS2,2S,5PreS2BPreS1T,L(S+PreS2+PreS1)Biotech2nologyGeneralBio2Hep2BPreSSCHOS2S6016211215g/,1100%-S,215gBio2Hep2B10gEngrix2B(SKB)7,HBcAg18518321kD,C,27nm,,,,HBcAgT,CTL,(SML)1987Clarke8(FMDV)HBcAgN,,HBcAg842001293©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(CHOC127CV21)HBsAg,,22nm,HBsAgCHOC12710CHOHBsAg,C127HBsAg,,,,,,HBsAg(23g/ml),;,,,,,Valenzuela11HBsAg(ADH-1)(Saccharomycescervisiae),(PHO),(PGK)HBsAgHBsAg22nm,,,SKB()MSD()(),,,40gHBsAg,,,,10HBsAg,(Hansenulapolymorpha,H.P.)12(Pichiapostoris,P.P.),,,,,,,,1P.pastorisH.polymorphaP.pastorisH.polymorphaAOXMOX,FMD//(100)His4Ura3,Leu2HARS,,,942001293©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.:MOX()FMD()DAS()P1P1AOX1(),,30%80%H1P1RhineHBsAg(adw),H1P1:30adw2HBsAg,H1P1,,,100g22nmHBsAg400g/ml,HBsAg60g/ml,,,99%,(BALB/c)(No.937CHONo.931101)4,,CHOC127HBeAg,:SDS2PAGE5HBsAg24kDP24,;CHOC127HBsAgP24,(GP27),25%13,CHOGP30,40%,GP30CHO2rHBsAg485,,14,(BALB/c),CHO15AbbottRIA016(10g),CHO(10g)(510g)GMT(mIU),2Table2ComparisonofHBsAbtitersinpupilsimmunizedwithHBvaccineofdifferentkindsVaccinesDose(g)No.oftested3712(Mo.)Serocon2version(%)GMTSerocon2version(%)GMTSerocon2version(%)GMT1-37(p)10308824.4796278.7992215.219203(p)103096150.69100365.59100457.34MSD(Y)5589321.5996259.2194115.73Amgen(Y)10579133.3796243.5994291.249312623(Y)5569330.2894155.4992.73110.359312645(Y)51068827.3490133.5588.54122.38921101(C)10456037.3496254.4494123.47921102(C)10426435.6093268.1595167.93Note:P:plasmavaccine,C:CHO2rDNAvaccine,Y:yeastrDNAvaccine052001293©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(C127)345Table3ThepreventiveefficacyagainstperinataltransmissiontoneomatesborntomotherspositiveforHBsAgPlasmavaccineDose(g)No.ofneonatesPreventiverate(%)at89mo.GMT(mIU/ml)at89mo.Ch2B85322207854.481.6Ch2L703222205560.0233.5Ch2B831404286.6359.6:(,1991,7)Table4ThepreventiveefficacyagainstperinataltransmissiontoneonatesborntomothrspositiveforHBsAgandHBeAg(CHOrDNAvaccine)CHOrDNAvaccineDose(g)No.ofneonatesPreventiverate(%)at89mo.GMT(mIU/ml)at89mo.C127(Canada)205580.0185.4T5155(Pasteur)202050.0155.6Ch28910205176.1125.9Ch2921101104568.0128.8Table5ThepreventiveefficacyagainstperinataltransmissiontoneomatesborntomotherspositiveforHBsAgandHBeAg(yeastrDNAvaccine)YestrDNAvaccineDose(g)No.ofneonatesPreventiverate(%)at89mo.GMT(mIU/ml)at89mo.Amgen1010085.6884.44MSD59678.9264.70103085.2396.96Cs2Ys2C108292.0120.73SK207273.3167.46Beijing9362355086.7262.45Beijing921262053896.766.6,SK,,,5g20g,,B152001293©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(SL,SL=151)S/L3(L3),CHOS/LMSD/B1A(Gen2HB2Vax),:H.P.-rHBsAgCHO-rHBsAg:H.P.-rHBsAgS,CHO2rHBsAgS,PreS1PreS2H.P.-rHBsAg,CHO-rHBsAg3kD6kDH.P.-rHBsAg1411,CHO-rHBsAg34rHBsAg,CHOH.P.-rHBsAgadw2,CHO-rHB2sAgayw,BALB/cCTL2,CHO-rHBsAgH.P.-rHBsAg,CTLH.P.-rHBsAg2BALB/cCTLFig.2AntibodyandCTLresponseofBALB/cmicetodifferentpreparationofHBVantigens(DiminskyD,ShirmbeckR,etal,1997,Vaccine,15:6372647)rHBsAg,H.P.S(1g)S(2g)S/L(2g)S/L3(2g))Gen2HB2Vax(2g)BALB/cCTL173,:H.P.-rHBsAg50ng1gCTLHB2sAg,CTL,,CTL;H.P.rHBsAgSL,SS/LS/L3252001293©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(%specificrelease)3BALB/cCTLFig.3AntibodyandCTLresponseBALB/cmicetodifferentpreparationofHBVSantigen(R.Schirmbeck,JVirolog,1994,68(3):141821425)DiminskyCHO2rHBsAgCTLPreSSCTL,,CTLH.P.HBsAg,,,,rHBsAg,,,,,,S(5%),,,,LHBcAg,,:1KrugmanS,GilesJP,HammondJ.HepatitisVirus:effectofheatontheinfectivityandantigenicityoftheMS21andMS22strainsJ.JInfectDis1970,122(5):4322436.2NeurathAR,KentSBH,StrickN.Detectionofantiviralanti2bodieswithpredeterminedspecificityusingsyntheticpeptide22lactamaseconjugated:applicationtoantibodiesspecificforthePresantigenofthehepatitisBvirusenvelopeproteinsJ.JGenVirol,1986,67:4532461.3MichelML,Po