重组血小板糖蛋白ibα活性片段在cho细胞中表达的论文

《重组血小板糖蛋白ibα活性片段在cho细胞中表达的论文》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、重组血小板糖蛋白Ibα活性片段在CHO细胞中表达的论文重组血小板糖蛋白Ibα活性片段在CHO细胞中表达的论文【摘要】目的：建立表达重组糖蛋白ibα活性片段（rgpibαh1v289）细胞株，并且纯化其表达产物，研究其生物学功能。方法：利用脂质体将含有编码gpibαh1v289的dna质粒pcmv3转染cho细胞。用亲和层析法纯化重组片段。用sdspage和westernblot鉴定纯化产品的纯度和免疫学活性。用血小板聚集法检测纯化产品对瑞斯托霉素诱导血小板聚集功能的影响。用elisa测定纯化产品对vw

2、f与血小板结合能力的影响。结果：每1l培养上清液可纯化到6.15mgrgpibαh1v289重组产品。sdspage显示，纯化的重组片段主要在42kd处显一蛋白区带，westernblot显示，抗gpibα单抗sz2能与重组片段在42kd处显单一蛋白区带。纯化重组片段能抑制瑞斯托霉素诱导的血小板聚集功能。elisa测定显示纯化产品抑制血浆vwf与血小板的结合能力。结论：cho细胞株a1能恒定表达rgpibαh1v289，重组片段具有较高的纯度、免疫学活性和生物学活性，有可能开发为有效的抗血栓药物。【

3、关键词】cho细胞重组糖蛋白ibα表达纯化studyofexpressioninchinesehamsterovarycellsandfunctionofrebinantactivefragmentofglycoproteinibαh1v289zhaoyiming,shaobojing,dongningzheng，shenfei，ruanchanggeng.jiangsuinstituteofhematology,thefirstaffiliatedhospitalofsoochowunivers

4、ity,suzhou215006,china［abstract］objective:tostudythebiologicalfunctionofglycoproteinproteinibα,acelllineexpressingrebinantactivefragmentrgpibαh1v289wasconstructedandpurifiedfragmentwasacquiredtoresearchitsfunction.methods:thechocellsweretransfectedwithpla

5、smidpcmv3includinggpibαh1v289dnabylipofectamine.therebinantfragmentwaspurifiedwithaffinitychromatographycolumn.thepurityandimmuneactivityofpurifiedproductswereidentifiedwithsdspageandwesternblotrespectively.theeffectofrgpibαh1v289ontheplateletaggregatio

6、nfunctionwasdetectedwithplateletaggregationassay.theeffectofpurifiedproductonbindingactivitybetweenplasmavwfandplateletwasmeasuredwithelisa.results:6.15mgrebinantfragmentinculturesupernatantsperlitercouldbepurifiedwithaffinitychromatography.therebinantfrag

7、mentformedamainlaneatthepositionof42kdwithsdspageandcouldreactwithmonoclonalantibodysz2bywesternblot.itcouldinhibittheplateletaggregationinducedbyristocetinandinhibitbindingactivitybetweenplasmavwfandplatelets.conclusion:chocelllinea1canexpressrgpibαh1v

8、28stablyandtherebinantfragmentwhichhashighpurity,immuneactivityandbiologyactivity,canbeapotentialantithrombosisdrug.［keywords］chinesehamsterovarycells；rebinantglycoproteinibα；express；purification在正常生理条件下，循环中