title: Expression and Characterization of a Single-Chain Variable Fragment against Human LOX-1 inEscherichia coliandBrevibacillus choshinensis.
abstract: The single-chain variable fragment (scFv) against lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a promising molecule for its potential use in the diagnosis and immunotherapy of atherosclerosis. Producing this scFv in several milligram amounts could be the starting point for further engineering and application of the scFv. In this study, the abundant expression of the anti-LOX-1 scFv was attempted usingEscherichia coli(E. coli) andBrevibacillus choshinensis (B. choshinensis). The scFv had limited soluble yield inE. coli, but it was efficiently secreted byB. choshinensis. The optimized fermentation was determined using the Plackett-Burman screening design and response surface methodology, under which the yield reached up to 1.5 g/l in a 5-L fermentor. Moreover, the properties of the scFvs obtained from the two expression systems were different. The antigen affinity, transition temperature, and particle diameter size were 1.01E-07 M, 55.2 ± 0.3°C, and 9.388 nm for the scFv expressed byB. choshinensis, and 4.53E-07 M, 52.5 ± 0.3°C, and 13.54 nm for the scFv expressed byE. coli. This study established an efficient scale-up production methodology for the anti-LOX-1 scFv, which will boost its use in LOX-1-based therapy.
authors.affiliation: School of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
authors.affiliation: School of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China.
authors.forename: Jun-Yan
authors.initials: JY
authors.lastname: Xiang
authors.affiliation: School of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China.
authors.forename: Ping
authors.initials: P
authors.lastname: Kong
authors.affiliation: School of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Liu
authors.affiliation: School of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China.|Key Laboratory for Molecular Enzymology and Engineering, Ministry of Education, Jilin University, Changchun, Jilin 130012, P.R. China.|National Engineering Laboratory for AIDS Vaccine, School of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China.
authors.forename: Qiuhong
authors.initials: Q
authors.lastname: Xie
authors.affiliation: School of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China.|Key Laboratory for Molecular Enzymology and Engineering, Ministry of Education, Jilin University, Changchun, Jilin 130012, P.R. China.|National Engineering Laboratory for AIDS Vaccine, School of Life Science, Jilin University, Changchun, Jilin 130012, P.R. China.
authors.forename: Hongyu
authors.initials: H
authors.lastname: Xiang
mesh_terms: D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D058675:Brevibacillus / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D006801:Humans; D055550:Protein Stability; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D051127:Scavenger Receptors, Class E / Q000187:drug effects*; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D013696:Temperature; D013997:Time Factors
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D051127:Scavenger Receptors, Class E; D057127:Single-Chain Antibodies
keywords: Brevibacillus choshinensis; Escherichia coli; Lectin-like oxidized low-density lipoprotein receptor-1; production methodology; single-chain variable fragment antibody