Onishi Hiromasa, Mizukami Makoto, Hanagata Hiroshi, Tokunaga Masao, Arakawa Tsutomu, Miyauchi Akira
Bio Research Group, R&D Department, Higeta Shoyu Co., Ltd., 2-8 Chuo-cho, Choshi, Chiba 288-8680, Japan.
Protein Expr Purif. 2013 Oct;91(2):184-91. doi: 10.1016/j.pep.2013.08.005. Epub 2013 Aug 20.
Expression of scFv in Brevibacillus choshinensis was tested using combinations of three different promoters and four different secretion signals. Two model scFv constructs, i.e., His-scFvFLU and His-scFvHEL, were successfully expressed with some of the combinations. Ni Sepharose column and size exclusion chromatography resulted in fairly pure preparations of these two proteins. The purified His-scFvFLU inhibited fluorescence from fluorescein, while the purified His-scFvHEL inhibited lysozyme activity. Relatively high yield of His-scFvFLU (∼40%) and His-scFvHEL (∼30%) was achieved with the expression and purification system described here.
使用三种不同启动子和四种不同分泌信号的组合，测试了单链抗体片段（scFv）在嗜碱芽孢杆菌中的表达。两种模型scFv构建体，即His-scFvFLU和His-scFvHEL，通过其中一些组合成功表达。镍琼脂糖柱和尺寸排阻色谱法得到了这两种蛋白质的相当纯的制剂。纯化的His-scFvFLU抑制了荧光素的荧光，而纯化的His-scFvHEL抑制了溶菌酶活性。使用本文所述的表达和纯化系统，His-scFvFLU（约40%）和His-scFvHEL（约30%）实现了相对较高的产量。
