Department of Bioengineering , University of Pennsylvania , Philadelphia , Pennsylvania 19104 , United States.
Bioconjug Chem. 2019 Jan 16;30(1):63-69. doi: 10.1021/acs.bioconjchem.8b00720. Epub 2018 Dec 26.
We demonstrate selective labeling of cell surface proteins using fluorogen-activating proteins (FAPs) conjugated to standard immunoglobulins (IgGs). Conjugation was achieved with a polypeptide reagent comprised of an N-terminal photoactivatable Fc-binding domain and a C-terminal FAP domain. The resulting FAP-antibody conjugates were effective agents for protein detection and cell ablation in cultured mammalian cells and for visualizing cell-cell contacts using a tethered fluorogen assay. Because our approach allows FAP-antibody conjugates to be generated for most currently available IgGs, it should have broad utility for experimental and therapeutic applications.
我们展示了使用与标准免疫球蛋白(IgG)缀合的荧光素酶激活蛋白(FAP)对细胞表面蛋白进行选择性标记。缀合是通过由 N 端光活化 Fc 结合结构域和 C 端 FAP 结构域组成的多肽试剂实现的。所得的 FAP-抗体缀合物是在培养的哺乳动物细胞中进行蛋白质检测和细胞消融以及使用连接的荧光素酶测定法可视化细胞-细胞接触的有效试剂。由于我们的方法允许为大多数现有的 IgG 生成 FAP-抗体缀合物,因此它应该在实验和治疗应用中具有广泛的用途。