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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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抗vegfr-2全人源igg1抗体的构建及其在cho-k细胞中的表达
, PP. 1544-1549
Keywords: vegfr-2,全长抗体,抗肿瘤血管生成
Abstract:
本文在实验室构建的单链抗体-fc融合抗体[scfv(ak404r)-fc]的基础上构建抗vegfr-2全人源igg1样全长抗体(mab-04)。利用重叠pcr,获得mab-04的轻链和重链的核酸序列后分别克隆到真核表达载体pcdna3.1,获得重组质粒。脂质体法将重组质粒转染至cho-k细胞,经proteina柱纯化细胞培养上清液获得目的蛋白,利用westernblotting检测目的蛋白,elisa检测mab-04与抗原亲和力。测序表明重组质粒构建成功,westernblotting检测显示目的蛋白成功表达(1μgsml-1),elisa检测阐明该抗体能与抗原结合并呈浓度依赖性(ic50为50nmolsl-1),表明mab-04成功表达并正确装配,为进一步大量制备该抗体及其活性研究打下基础。
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