資源描述：
《鼠粒細(xì)胞巨噬細(xì)胞集落刺激因子的純化和復(fù)性研究.pdf》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。
1、藥物生物技術(shù)386PharmaceuticalBiotechnology2013，20(5)：386～391PurificationandRenaturationofE．coli．DerivedMouseGranulocyte-MacrophageColonyStimulatingFactorCAORong．yue，CHENMeng，GEChi—yu，TANGXiao—tian，LIFei，ZHANGPeng，XUMao—lei，lLIUJing-jingf《1．MinigenePharmacyLaboratory,Sch
2、oolofLifeScienceandTechnology,ChinaPharmaceuticalUniversity，Nan—jg210009China；2．XintrumPharmaceuticalsLtd,N鋤ng210017,China)AbstractGranulocytemacrophage—colonystimulatingfactor(GM·CSF)isahematopoieticgrowthfactorandaproinflammatorycyt0kine．RecombinantmouseGM．CSFfr
3、mGM—CSF)isnecessaryinantineoplasticandanti—inflammatoryresearch．ToobtainrmGM—CSF．theencodingregionformGM．CSFwasclonedbetweenNcoIand口0mHIinpET28a．TheproteinwasexpressedintheformofinclusionbodiesinEc0lihoststrainBL21fDE3)．Theexpressionlevelwasmorethan51．6％oftotalcel
4、llysate．Inclusionbodieswerewashedunderoptimizedconditions，anddissolvedinabufercontaining8mol／Lurea．Dilutionmethodcombinedwithpolyethy—leneglyeolfPEG1asaco．solventwaSemployedtorefoldproteinsin2mol／Lurea．Afterion—exchangechromatography，rmGM—CSFwithahighDuritywasobta
5、ined．PurifiedrmGM—CSFhasagoodfidelity，whichmakesitpossibletopromoteproliferationofmyeloidce11sinitroandtoincreasethenumberofleukocyteandneutrophilinvivo．Ahighyieldof8．06mgrmGM—CSFperlitreofcellculturewasachievedbyusingthismethod．Owingtoitshighyieldandsimpleoperati
6、on，thisprotocolissuitableforlarge—scaleproduction·KeywordsRecombinantmousegranulocyte-macrophagecolony—stimulatingfactor，Inclusionbodies，Polyethyleneglycol，Purifi—cation，Refolding，BioactivityCLCNumberQ813DocumentcodeAArticleID1005—8915(2013)05—0386—06agreatdealofi
7、solationtrials_9jGranulocytemacrophage-colonystimulatingfactor(GM—．whichrevealedconundrumofCSF)wasoriginallydugoutasapotentialofgeneratingbothhighcostsandlowyield，thoughglycosylatedproteinwasgranulocyteandmacrophagecoloniesfromhematopoieticcellsobtained．However，th
8、ebiologicalactivityofmouseGM—CSFinmousebonemarrow”．However．a(chǎn)seriesoffunctionsweredoesnotdependonglycosylationstiesbutcorrectsconforma—discoveredsubseque
