title: Efficient production of anti-fluorescein and anti-lysozyme as single-chain anti-body fragments (scFv) by Brevibacillus expression system.
abstract: 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.
authors: Bio Research Group, R&D Department, Higeta Shoyu Co., Ltd., 2-8 Chuo-cho, Choshi, Chiba 288-8680, Japan.
authors: Hiromasa
authors: H
authors: Onishi
authors: Makoto
authors: M
authors: Mizukami
authors: Hiroshi
authors: H
authors: Hanagata
authors: Masao
authors: M
authors: Tokunaga
authors: Tsutomu
authors: T
authors: Arakawa
authors: Akira
authors: A
authors: Miyauchi
mesh_terms: D058675:Brevibacillus / Q000235:genetics* / Q000378:metabolism; D002850:Chromatography, Gel; D019793:Fluorescein / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D009113:Muramidase / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D009113:Muramidase; D019793:Fluorescein
keywords: Brevibacillus choshinensis; Expression system; Recombinant protein; Secretory expression; Signal peptide; Single-chain antibody (scFv)