抗β淀粉样肽40人单克隆抗体在昆虫细胞分泌表达.pdfVIP

J Med No．6 ·论 善· Res，Jun2007，V01．36 抗B淀粉样肽40人单克隆抗体在 昆虫细胞分泌表达 蔡炯 郭 瑜 梁米芳 李 方 刘 飞 王世真 摘要 目的通过杆状病毒昆虫细胞系统重组表达抗8淀粉样肽40人单克隆抗体，为全人抗体治疗阿尔茨海默病创造 条件。方法 病毒的昆虫细胞培养液上清过蛋白A一琼脂糖亲和层析柱，洗脱纯化出的抗体经过SDS—PAGE电泳鉴定纯化蛋白质。结果 胞后可见昆虫细胞表达人抗体。经过蛋白A一琼脂糖亲和层析，从昆虫细胞培养基中纯化出抗B淀粉样肽40人单克隆抗体。 结论通过基因工程手段在人抗A1340抗体可变区羧基端连接人抗体恒定区片段，通过杆状病毒昆虫系统分泌表达并纯化获得 完整的人抗A1340单克隆抗体。 关键词 阿尔茨海默病 B淀粉样肽 全人抗体 昆虫细胞 Secreted ofHumanMonoclonal inInsectCells．Cai Expression AntibodyAgainstAmyloidBPeptide Jiong，GuoYu，LiangMifang，Li Nuclear and 100730，China Fang，LiuFei，WangShizhen．Departmentof medicine，PUMCHospital，CAMSPUMC，Beijing Abstract To B 40humanmonoclonalinbaculovirusinsect asto Objectiveexpressanti—amyloidpeptide antibody system，SO forAlzheimer’S waseonstruc— acandidate disease provide antibodytherapeutics patients．MethodspAC—VH—VLexpressionpalsmid ted and chainvariable into MCS．The wereco—infectedwith heavy palsmids bycloning light regionseparatelypAC—L—CH3 expression BaculoGoldvirusintoSF9cellsandthe wasdetectedimmunoflureseencemethod．Theculture of IgGexpression by supernatant ～infectedcellswasloadedon A andtheeluted was toSDS—PAGE pre—equilibratedproteinSepharose antibodysubjected suitsThe wasconstructedas infectedinsectcellssecretedhuman afterCO— expressionplasmidpAC—VH—VL expected．The antibody infectedwith and B 40humanmonoclonalwas fromcultureSU—