Chen Qi, Rozovsky Sharon, Chen Wilfred
Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE 19716, USA.
Chem Commun (Camb). 2017 Jul 4;53(54):7569-7572. doi: 10.1039/c7cc04246a.
Outer membrane vesicles (OMVs) are proteoliposomes derived from the outer membrane and periplasmic space of many Gram-negative bacteria including E. coli as part of their natural growth cycle. Inspired by the natural ability of E. coli to sort proteins to both the exterior and interior of OMVs, we reported here a one-pot synthesis approach to engineer multi-functionalized OMV-based sensors for both antigen binding and signal generation. SlyB, a native lipoprotein, was used a fusion partner to package nanoluciferase (Nluc) within OMVs, while a previously developed INP-Scaf3 surface scaffold was fused to the Z-domain for antibody recruiting. The multi-functionalized OMVs were used for thrombin detection with a detection limit of 0.5 nM, comparable to other detection methods. Using the cohesin domains inserted between the Z-domain and INP, these engineered OMVs were further functionalized with a dockerin-tagged GFP for cancer cell imaging.
外膜囊泡（OMVs）是源自包括大肠杆菌在内的许多革兰氏阴性菌外膜和周质空间的蛋白脂质体，是其天然生长周期的一部分。受大肠杆菌将蛋白质分选到OMVs外部和内部的天然能力启发，我们在此报告了一种一锅法合成方法，用于设计基于OMV的多功能传感器，用于抗原结合和信号产生。天然脂蛋白SlyB用作融合伙伴，将纳米荧光素酶（Nluc）包装在OMVs内，而先前开发的INP-Scaf3表面支架与Z结构域融合以招募抗体。多功能化的OMVs用于凝血酶检测，检测限为0.5 nM，与其他检测方法相当。利用插入Z结构域和INP之间的黏连蛋白结构域，这些工程化的OMVs进一步用带有dockerin标签的绿色荧光蛋白（GFP）进行功能化，用于癌细胞成像。
