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链间连接肽对单链双特异性抗体生物学活性的影响
  • 摘要

    单链双特异性抗体(scBsAb)是具有良好临床应用前景的基因工程抗体, 但不同的链间连接肽(interlinker)对其生物学活性的影响尚有待深入研究.设计并合成3种不同的链间连接肽序列, 分别命名为Fc, HSA和205C′, 并构建成单链双特异性抗体通用表达载体, 以抗人CD3改形单链抗体(scFv)和抗人卵巢癌单链抗体为基础, 构建成单链双特异性抗体.SDS-PAGE及Western blot结果表明, 3种不同的链间连接肽对抗体的表达量无明显影响.在此基础上, 进行ELISA及血液药代动力学测定, 发现不同链间连接肽的单链双特异性抗体与相应抗原的结合活性及其在小鼠体内的清除相T1/2存在一定差异, 链间连接肽HSA可以显著延长抗体在体内的滞留时间.上述研究结果提示, 链间连接肽序列将会影响单链双特异性抗体的抗原结合活性及其在体内的稳定性等重要的生物学特性.因此, 筛选理想的链间连接肽序列对于构建单链双特异性抗体具有重要意义.合适的链间连接肽可以赋予单链双特异性抗体更适于临床应用需求的生物学活性.

  • 作者

    方敏  蒋欣  杨志  俞小淙  尹长城  李骅  赵瑞  张众  林晴  黄华樑 

  • 作者单位

    中国科学院遗传与发育生物学研究所,北京,100101/北京大学临床肿瘤学院,北京肿瘤医院,北京,100036

  • 刊期

    2003年18期 ISTIC PKU

  • 关键词

    单链抗体  双特异性抗体  单链双特异性抗体  链间连接肽 

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