Veterinary Research Institute, Hudcova 296/70, 621 00, Brno, Czech Republic.
Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 753/5, 625 00, Brno, Czech Republic.
Sci Rep. 2017 Dec 13;7(1):17501. doi: 10.1038/s41598-017-17951-5.
MS2 phage-like particles (MS2 PLP) are artificially constructed pseudo-viral particles derived from bacteriophage MS2. They are able to carry a specific single stranded RNA (ssRNA) sequence of choice inside their capsid, thus protecting it against the effects of ubiquitous nucleases. Such particles are able to mimic ssRNA viruses and, thus, may serve as the process control for molecular detection and quantification of such agents in several kinds of matrices, vaccines and vaccine candidates, drug delivery systems, and systems for the display of immunologically active peptides or nanomachines. Currently, there are several different in vivo plasmid-driven packaging systems for production of MS2 PLP. In order to combine all the advantages of the available systems and to upgrade and simplify the production and purification of MS2 PLP, a one-plasmid double-expression His-tag system was designed. The described system utilizes a unique fusion insertional mutation enabling purification of particles using His-tag affinity. Using this new production system, highly pure MS2 PLP can be quickly produced and purified by a fast performance liquid chromatography (FPLC) approach. The system can be easily adapted to produce other MS2 PLP with different properties.
MS2 噬菌体样颗粒（MS2 PLP）是人工构建的源自噬菌体 MS2 的假病毒颗粒。它们能够在衣壳内携带特定的单链 RNA（ssRNA）序列，从而保护其免受普遍存在的核酸酶的影响。这些颗粒能够模拟 ssRNA 病毒，因此可以作为分子检测和定量此类制剂的过程控制，这些制剂存在于多种基质、疫苗和疫苗候选物、药物传递系统以及免疫活性肽或纳米机器的展示系统中。目前，有几种不同的体内质粒驱动包装系统可用于生产 MS2 PLP。为了结合现有系统的所有优势，并升级和简化 MS2 PLP 的生产和纯化，设计了一种单质粒双表达 His 标签系统。该系统利用独特的融合插入突变，可使用 His 标签亲和性进行颗粒的纯化。使用这种新的生产系统，可以通过快速高效液相色谱（FPLC）方法快速生产和纯化高纯度的 MS2 PLP。该系统可以轻松适应生产具有不同特性的其他 MS2 PLP。
