• 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

        Assays for Selection of Single-Chain Fragment Variable Recombinant Antibodies to Metal Nanoclusters

        互联网

        905
        The protocols herein describe colony-lift and fluorescent immunoassays that were used to identify bacterial colonies that produced single-chain fragment variable (ScFv) recombinant antibodies reactive with zero-state silver. A large (approx 2.9-billion member) phage-displayed antibody library was panned against zero valent silver. Bacterial colonies obtained after two rounds of selection were either lifted onto nitrocellulose filters or picked to individual wells of 384-well microtiter culture plates. Colonies lifted onto filters were placed onto zero valent silvercoated filters and induced to produce soluble ScFv antibodies. ScFv antibodies, expressed by individual colonies, that bound to silver nanocluster-coated filters were detected using an anti-ScFv antibody conjugated to horseradish peroxidase and a chemiluminescent substrate. Colonies picked to 384-well micotiter culture plates were induced to express soluble ScFv antibodies. ScFv antibodies in bacterial periplasmic extract were transferred from 384-well culture plates to 384-well assay plates containing zero-state silver particles and an anti-ScFv antibody conjugated to a fluorescent dye. ScFv antibodies, expressed by individual bacterial clones, that bound to zero valent silver nanoparticles in 384-well assay plates were detected using an FMAT 8100 fluorescent plate reader. The colony-lift and fluorescent immunoassays detected ScFv antibodies reactive with zero valent silver. Similar assay formats should also be useful to detect bacterially expressed recombinant antibodies or proteins to other nanoclusters.
        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