Hu Wei, Xiang Jun-Yan, Kong Ping, Liu Ling, Xie Qiuhong, Xiang Hongyu
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.
J Microbiol Biotechnol. 2017 May 28;27(5):965-974. doi: 10.4014/jmb.1702.02007.
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 using () and . The scFv had limited soluble yield in , but it was efficiently secreted by . 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 by , and 4.53E-07 M, 52.5 ± 0.3°C, and 13.54 nm for the scFv expressed by . 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.
抗凝集素样氧化低密度脂蛋白受体-1（LOX-1）的单链可变片段（scFv）因其在动脉粥样硬化诊断和免疫治疗中的潜在应用而成为一种有前景的分子。以数毫克的量生产这种scFv可能是其进一步工程化和应用的起点。在本研究中，尝试使用（）和（）来大量表达抗LOX-1 scFv。该scFv在（）中的可溶性产量有限，但它能被（）有效分泌。使用Plackett-Burman筛选设计和响应面方法确定了优化的发酵条件，在此条件下，在5升发酵罐中产量可达1.5克/升。此外，从两种表达系统获得的scFv的性质不同。由（）表达的scFv的抗原亲和力、转变温度和粒径分别为1.01E-07 M、55.2±0.3°C和9.388纳米，由（）表达的scFv的抗原亲和力、转变温度和粒径分别为4.53E-07 M、52.5±0.3°C和13.54纳米。本研究建立了一种高效的抗LOX-1 scFv放大生产方法，这将促进其在基于LOX-1的治疗中的应用。
