中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2010年
2期
113-115
,共3页
邱丰%贾志远%郭敏卓%陈斯勇%伊瑶%沈立萍%余陶%费永亮%郭瑜%毕胜利
邱豐%賈誌遠%郭敏卓%陳斯勇%伊瑤%瀋立萍%餘陶%費永亮%郭瑜%畢勝利
구봉%가지원%곽민탁%진사용%이요%침립평%여도%비영량%곽유%필성리
肝炎病毒%病毒融合蛋白质类%疫苗%免疫测定
肝炎病毒%病毒融閤蛋白質類%疫苗%免疫測定
간염병독%병독융합단백질류%역묘%면역측정
Hepacivirus%Viral fusion proteins%Vaccines%Immunassay
目的 获得高纯度的丙型肝炎亚单位融合蛋白并评价其免疫原性.方法 利用原核表达系统,以pET-11d作为载体,在大肠埃希菌BL21(DE3)中经IPTG诱导表达获得融合蛋白,之后采用DEAE阴离子交换和镍离子柱亲和层析进行纯化.通过Western Blot验证融合蛋白的抗原活性.同时以该蛋白免疫实验动物后测定血清中的抗体滴度.结果 成功获得很高纯度的丙型肝炎哑单位融合蛋白,EIA显示融合蛋白能够诱生出高滴度的抗体.结论 原核表达系统表达的丙型肝炎病毒亚单位融合蛋白具有较强的免疫原性,为丙型肝炎病毒治疗性和预防性疫苗的研制提供了一条新的途径.
目的 穫得高純度的丙型肝炎亞單位融閤蛋白併評價其免疫原性.方法 利用原覈錶達繫統,以pET-11d作為載體,在大腸埃希菌BL21(DE3)中經IPTG誘導錶達穫得融閤蛋白,之後採用DEAE陰離子交換和鎳離子柱親和層析進行純化.通過Western Blot驗證融閤蛋白的抗原活性.同時以該蛋白免疫實驗動物後測定血清中的抗體滴度.結果 成功穫得很高純度的丙型肝炎啞單位融閤蛋白,EIA顯示融閤蛋白能夠誘生齣高滴度的抗體.結論 原覈錶達繫統錶達的丙型肝炎病毒亞單位融閤蛋白具有較彊的免疫原性,為丙型肝炎病毒治療性和預防性疫苗的研製提供瞭一條新的途徑.
목적 획득고순도적병형간염아단위융합단백병평개기면역원성.방법 이용원핵표체계통,이pET-11d작위재체,재대장애희균BL21(DE3)중경IPTG유도표체획득융합단백,지후채용DEAE음리자교환화얼리자주친화층석진행순화.통과Western Blot험증융합단백적항원활성.동시이해단백면역실험동물후측정혈청중적항체적도.결과 성공획득흔고순도적병형간염아단위융합단백,EIA현시융합단백능구유생출고적도적항체.결론 원핵표체계통표체적병형간염병독아단위융합단백구유교강적면역원성,위병형간염병독치료성화예방성역묘적연제제공료일조신적도경.
Objective Obtain the hepatitis C virus high purified subunit fusion protein and detect its immunogenicity.Methods With the vector of pET-11d,fusion protein was expressed in Escherichia coli BL21(DE3) after induced by IPTG.The protein was then purified by DEAE negative ion exchange chromatography and Ni2+ affinity chromatography.Western Blot analysis was used to detect the antigenicity of the fusion protein.At the same time,the sera were collected and prepared from the immunized experimental animals in order to investigate the immunogenicity of the protein by EIA.Results High purified hepatitis C virus subunit fusion protein was obtained successfully.The EIA indicated that the fusion protein could elicit specific antibodies in the animals with very high titers.Conclusion The hepatitis C virus subunit fusion protein expressed in prokaryotic system was proved to have strong immunogenicity.It could provide some helpful and useful information to the hepatitis C virus prophylactic and therapeutic vaccine development.
