Biotechnology, Sigma-tau, SpA, Pomezia (RM) 00040, Italy.
BMC Biotechnol. 2013 Sep 30;13:79. doi: 10.1186/1472-6750-13-79.
Consistent progress in the development of bacteriophage lambda display platform as an alternative to filamentous phage display system was achieved in the recent years. The lambda phage has been engineered to display efficiently multiple copies of peptides or even large protein domains providing a powerful tool for screening libraries of peptides, proteins and cDNA.
In the present work we describe an original method for dual display of large proteins on the surface of lambda particles. An anti-CEA single-chain antibody fragment and green fluorescent protein or alkaline phosphatase were simultaneously displayed by engineering both gpD and gpV lambda proteins.
Here we show that such modified phage particles can be used for the detection of target molecules in vitro and in vivo. Dual expression of functional moieties on the surface of the lambda phage might open the way to generation of a new class of diagnostic and therapeutic targeted nanoparticles.
近年来,噬菌体 λ 展示平台作为丝状噬菌体展示系统的替代方案,在发展上取得了持续的进展。λ噬菌体经过工程改造,可以高效展示多个肽段甚至是大的蛋白质结构域,为筛选肽段、蛋白质和 cDNA 文库提供了强大的工具。
在本工作中,我们描述了一种在 λ 噬菌体表面双重展示大蛋白的原创方法。通过工程化 gpD 和 gpV λ 蛋白,同时展示了抗 CEA 单链抗体片段和绿色荧光蛋白或碱性磷酸酶。
我们的研究结果表明,这种经过修饰的噬菌体颗粒可用于体外和体内靶分子的检测。λ噬菌体表面功能性结构域的双重表达可能为开发新型诊断和治疗靶向纳米颗粒开辟道路。