D V Shanshin, V S Nesmeyanova, E V Protopopova, A A Shelemba, V B Loktev, D N Shcherbakov
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引用次数: 0
摘要
我们获得了能与黄病毒表面糖蛋白 E 特异性相互作用的全长人源化嵌合抗体 10H10ch。为了构建该抗体,我们使用了构成抗体活性中心的单克隆抗体 10H10 重链和轻链的可变片段,以及人类 IgG1 抗体重链恒定部分的片段。由此产生的全长嵌合人源化抗体 10H10ch 能与对人类致病的黄病毒(如蜱传脑炎、寨卡病毒、西尼罗河病毒和登革热病毒)的 E 蛋白发生特异性相互作用。通过 ELISA 和生物层干涉测量法对 10H10ch 抗体与一系列原生和重组黄病毒抗原的相互作用常数进行的免疫化学评估表明,嵌合抗体的解离常数(Kd)在纳摩尔区,与高亲和力小鼠单克隆抗体 10H10 的解离常数相当。本文讨论了将所制备的嵌合人源化抗体 10H10ch 用于诊断、预防和治疗各种黄病毒感染的可能性。
Preparation and Construction of Chimeric Humanized Broadly Reactive Antibody 10H10 to Protein E of Tick-Borne Encephalitis Virus.
A full-length humanized chimeric antibody 10H10ch that specifically interacts with the surface glycoprotein E of flaviviruses was obtained. To construct it, we used variable fragments of the heavy and light chains of the monoclonal antibody 10H10 that form the active center of the antibody and a fragment of the constant part of the heavy chain of the human IgG1 antibody. The resulting full-length chimeric humanized antibody 10H10ch specifically interacted with the E protein of flaviviruses pathogenic to humans, such as tick-borne encephalitis, Zika, West Nile, and dengue viruses. An immunochemical assessment of the interaction constants of the 10H10ch antibody with a panel of native and recombinant flavivirus antigens by ELISA and biolayer interferometry showed that the dissociation constant (Kd) of the chimeric antibody is in the nanomolar region and is comparable to that of the high-affinity mouse monoclonal antibody 10H10. The possibility of using the resulting chimeric humanized antibody 10H10ch for the diagnosis, prevention, and treatment of various flavivirus infections is discussed.
期刊介绍:
Bulletin of Experimental Biology and Medicine presents original peer reviewed research papers and brief reports on priority new research results in physiology, biochemistry, biophysics, pharmacology, immunology, microbiology, genetics, oncology, etc. Novel trends in science are covered in new sections of the journal - Biogerontology and Human Ecology - that first appeared in 2005.
World scientific interest in stem cells prompted inclusion into Bulletin of Experimental Biology and Medicine a quarterly scientific journal Cell Technologies in Biology and Medicine (a new Russian Academy of Medical Sciences publication since 2005). It publishes only original papers from the leading research institutions on molecular biology of stem and progenitor cells, stem cell as the basis of gene therapy, molecular language of cell-to-cell communication, cytokines, chemokines, growth and other factors, pilot projects on clinical use of stem and progenitor cells.
The Russian Volume Year is published in English from April.