脂多糖
脂多糖结合蛋白
等温滴定量热法
重组DNA
细菌外膜
化学
细菌
大肠杆菌
产量(工程)
体外
单克隆抗体
生物化学
抗体
生物
免疫学
材料科学
基因
受体
冶金
CD14型
遗传学
作者
Jie Qiao,Ce Dong,Xinping Wang,Yi Liu,Lixin Ma
标识
DOI:10.1016/j.pep.2019.01.008
摘要
Human lipopolysaccharide (LPS) binding protein (LBP) is a ∼60 kDa glycosylated protein that mediates potent innate immune against invading Gram-negative bacteria by recognition of LPS in their outer membranes. To date, there is no method for efficient production of bioactive LPS-free LBP at sufficient amounts through prokaryotic expression system. Here we present a simple approach for rapid preparation of human LBP from a LPS-eliminated E. coli strain named ClearColi BL21 (DE)3. Combined with the usage of an ultra-high-affinity CL7/Im7 purification system, we achieved one-step purification of recombinant human LBP with over 90% purity at a yield of ∼4 mg/L when using LB culture medium. The produced LBP retains full LPS binding activity which was validated by fluorescence spectroscopy and isothermal titration calorimetry (ITC). Collectively, we develop a valid method that can be applied to cost-effectively produce and purify LPS-free proteins.
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