30422.新型疏水電荷誘導色譜介質hasepharose中蛋白質吸附性能研究

1、天津大學碩士學位論文新型疏水電荷誘導色譜介質HASepharose中蛋白質吸附性能研究姓名：金乃君申請學位級別：碩士專業(yè)：生物化工指導教師：白姝20090801ABSTRACTHydrophobicchargeinductionchromatography(HCIC)isanovelchromatographictechnique，inwhichaZOcyclicchemicalswereusedascandidatesofchroma

2、tographicligandInsuchcondition，bindingofproteinwasbasedonmoderatelyhydrophobicinteractionbetweenproteinandHCICligandwhiledesorptionwasachievedbytherepulsioninteractionbetweenionisableligandandproteinGenerallythepurificat

3、ionofproteinscouldbeeasilyobtainedwithoutcomplicatedpretreatmentsandharshelutionconditionThus，posttreatmentslikedesaltalsocouldbeofnoneedandthecostofpurificationwascutdownsignificantlyPreviouslyHASepharose，anoveladsorben

4、tforHCIC，wasdevelopedinourgroupbycouplinghistamineontoepoxyactivatedSepharoseCL一6BIncurrentthesis，adsorptionbehaviorofmodelproteins(1ysozyme，immunoglobulinGandbovineserumalbumin)onHASepharosewasinvestigatedextensivelyand

5、theinfluenceofionicstrengthandbufferpHonproteinadsorptionwasmeasuredFirstly，agarosegelSepharoseCL一6BwasemployedasmediumandactivatedwithepoxychloropropanebyintroducinghydrophilicsolventdimethylsulphoneWhentheconcentration

6、ofNaOH，theexperimentingtemperatureandtheactivatingtimewere08mol／150℃and3hoursrespectivelyactivatingdensityofepoxygroupscouldreached150Itmol／mlandaboveSecondly’toensureafairlyhighcouplingliganddensitythereactionwascarried

7、outataconcentrationoffreehistamine5timeshigherthanepoxydensityofactivatedmediumByextendingcouplingtime，increasingreactiontemperatureorpHtoanextent，ligandcouplingdensitycouldalsobeenhancedHASepharosewaspreparedWhentheconc

8、entrationoffreehistaminewas5timeshigherthanepoxydensityofactivatedmedium，withtheexperimentingtemperature60“(2andtheactivatingtime12hours，thedensityofligandcouldreachedabout150Bmol／m1Finallytheadsorptionsoflysozyme，bovine

9、serumalbuminandimmunoglobulinGonHA—SepharosewereobservedrespectivelyatdifferentionicstrengthsandpHExperimentalresultsshowedthat，whentheionicstrengthwassmallerthan15mol／1，theadsorptioncapacitiesofproteinsmerelychangedalit