• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构:
    • 成为企业机构
    • 个人用户:
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        Periplasmic Expression and Purification of Recombinant Fabs

        互联网

        975
        The ability to refine the affinity, specificity, and immunogenicity of recombinant antibodies (Abs) offers distinct advantages for the preparation of in vivo diagnostic and therapeutic immunoreagents. The bacterial expression vector pComb3 allows production of recombinant Ab Fabs (rFab) (1 ). Light-chain (LC) and heavy-chain (HC) Ab cDNA fragments are inserted separately into this vector, which produces heterodimeric rFab Abs that are isolated in a native form. Fabs generated with the pComb3 expression vector are documented to bind Ag with affinities similar to that of the parental hybridoma-generated Fab (2 ). In the plasmid, both chains are independently controlled by isopropyl-1-β-D-thioglactopyranoside (IPTG)-inducible lac expression. The original pComb3 vector allows for the display of rFabs on the surface of M13 filamentous phage and can be converted to produce rFabs in a soluble form. We have modified this vector to allow the expression and purification of soluble Fab Ab fragments using immobilized metal-affinity chromatography via a pentahistidine (His) tag fused to the HC constant region 1 Fig. 1 ; 2 ). A similar modification to pComb3 (pComb3x) designed exclusively for the expression of soluble rFabs has been performed by Barbas at the Scripps Research Institute (see Note 1 ).
        ad image
        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序