小鼠腹水IgG类单克隆抗体纯化方法的研究

:20051123:(30230620):(1969),,,.:(1950),,,,.IgG1,2,2,3,1,43(1.,130062;2.,130062;3.,130062;4.,130062):S865.1:A:10047034(2006)10001403:IgG;;;:(CAAS)(AS)DEAEIgG:CAASIgG6715%,AS(4312%),DEAE(100%);CAASIgG5112%,DEAE(819%),AS(6318%)IgGELISA:DEAEOD,DEAEIgGCAASIgG(McAb),,McAbMcAbMcAb,,McAb,,McAb(CAAS)(AS)DEAEIgGMcAb,SDSPAGEELISA3IgG11.1CS9000DUALWAVELENGTHFLYINGSPOTSCANNER,SHMADZU;DYY6B,;CNUV/WL;(TECAN)IgG,4E2,;SP2/0,(Caprylicacid),,;(Ammoniumsulfate),;DEAE,Sephan2dexG25phamacia;IgGHRP,1.21.2.14(60mmol/L,pH410),011mmol/LpH415(25L/mL),,30min,8000r/min30min,10110PBS,011mmol/LpH714(01227g/mL)30min,6000r/min15min,,PBS(1/5)100PBS20[1],41.2.2PBS,,,1h4,1000r/min10min,,PBS(1/5)100PBS420[2]1.2.3DEAESephandexG25,DEAE,0102mol/LNH4Ac(pH615),20[3]1.2.4,280nm260nmA280A260:/(mgmL-1)=(1145A280-0174A260)41HeilongjiangAnimalScienceandVeterinaryMedicine102006©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[4]1.2.5McAbSDSPAGE[5]0104%6167%4%SDS10%2212%115L60V,,120V,20%30min0125gR250250mL,,5,2h,,UVI,IgGMcAbMcAbMcAb1.2.6McAbELISAMcAb,SDSPAGE,IgG,,IgGSP2/0,OD2,113mgmL-12610755175110621971178610401741125312%SDSPAGE(1),McAb2,2,,IgG,50ku,IgG,25ku,,,,AS,CAAS,DE2AE1.,015L(26107mg/mL);2.,115L(1106mg/mL);3.,115L(1178mg/mL);4.,115L(0194mg/mL);5.,015L(55175mg/mL);6.,115L(2197mg/mL);71,115L(6104mg/mL);8.,115L(1145mg/mL);9.Marker1SDSPAGE,3,IgG2DEAEIgG100%,10%CAASIgGAS,DEAE;CAASIgGDEAE,ASELISAOD3DEAEOD,CAASASOD,DEAEIgG,IgGCAASAS,23IgG%IgG6715691243124411100100IgG51125019631863148198183ELISAOD49001110101018101010180011201830123014101053,DEAE100%,IgG,IgGASIgG,McAbCAAS51200610©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[1],,,.[J].,1995,11(2):5254.[2],.[M].:,2002:267.[3],,.[J].:,2002,31(1):108200.[4],,,.McAb/[J].,1999,30(4):455456.[5],DW.[M].:,2002:17131720.(009)ThestudyofpurifiedmethodsofmiceascitesIgGMcAbZHOUYu,LIYansong,PANFengguang,etal(JilinUniversity,Changchun130062,China)Keywords:MiceascitesIgG;caprylic;ammoniumsulfate;ionexchangechromatographyAbstract:MiceascitesIgGMcAbwaspurifiedbycaprylic/ammoniumsulfateprecipitation(CAAS),ammoniumsulfateprecipitation(AS)andionex2changechromatography(DEAE).ThepurityofIgGfromthemethodofCAAS(6715%)washigherthanthosefromAS(4312%)andlowerthanthosefromDEAE(100%).TherecoveryofIgGfromCAAS(5112%)washigherthanthosefromDEAE(819%)remarkablyandlowerthanthosefromAS(6318%)slightly.TheconcentrationofpurifiedIgGfromdifferentmethodswereadjustedthesameasthoseofunpurifiedascites.ELISAshowsthattheODvaluesofDEAEmethodwaslowerremarkable,itindicatedthattheimmunityofIgGwasdecreasedbythepurifiedmethodofDEAE.Conclusion:CAASisonesimple,lowcost,andefficientmethodforpurificationofIgGfrommiceascites.(009):20060110:(1963),(),,.(,133400):S816.8:A:10047034(2006)10001605:TMR;;;;:TMR,(P0105)6pH3NH3N(P0105),06(P0101),9h(P0101),9h/(P0101)CMCase,9hxylanase(P0105),6,69,(P0105)(P0101)(TotalMixedRation,TMR)20506020802090TMRTMRTMR,,,TMR,,TMR,TMR,TMR161HeilongjiangAnimalScienceandVeterinaryMedicine102006©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.