针对B细胞恶性瘤的抗CD22 scFv的表达和功能特征
Monireh Gholizadeh1,2, Shahriyar Abdoli3, Shafieeh Mansoori2
1Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Research in pharmaceutical sciences
|July 21, 2025
概括
这项研究优化了大肠杆菌中抗CD22单链可变片段 (scFvs) 的表达和净化. 混合净化产生了明显更容易溶解的蛋白质,具有强大的抗增殖作用,用于癌症治疗.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 免疫治疗是一种免疫疗法.
背景情况:
- 单链可变片段 (scFvs) 在癌症治疗中比单克隆抗体具有优势.
- 大肠杆菌 (E. coli) 的表达可以导致包容体的形成,阻碍治疗性蛋白质的产生.
- 优化scFv表达和净化对于开发有效的癌症治疗非常重要.
研究的目的:
- 在大肠杆菌中优化抗CD22 scFv (CD22-scFv) 的表达和净化.
- 为了评估纯化CD22-scFv的功能性质.
- 建立一个可扩展和具有成本效益的战略,生产功能性scFvs.
主要方法:
- CD22-scFv在大肠杆菌菌株Rosetta (DE3) 和Rosetta-gami 2中得到表达.
- 两种净化方法进行了比较:混合 (在柱子上重新折叠) 和本地条件.
- 蛋白质结合和抗增殖效应使用流动细胞计和MTT试验进行了评估.
主要成果:
- 在罗塞塔 (DE3) 中成功表达了CD22-scFv.
- 混合净化产生了15.86毫克/升,明显高于原生净化 (3.65毫克/升).
- 这两种纯化形式都与CD22+拉吉细胞结合;本地纯化的scFv表现出显著的抗增殖作用.
结论:
- 建立了一个可扩展和具有成本效益的方法来产生功能性的CD22-scFv.
- 优化的CD22-scFv表现出高特异性和抗增殖活性.
- 针对性治疗和诊断需要进一步的体内和临床研究.
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