中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (8): 851-856.doi: 10.4103/1673-5374.131611

• 原著:退行性病与再生 • 上一篇    下一篇

单链可变区片段融合蛋白可治疗重症肌无力

  

  • 收稿日期:2014-03-01 出版日期:2014-04-25 发布日期:2014-04-25
  • 基金资助:

    中国国家自然科学基金项目(30360100,30760234,30860260,81160373,81360458)

Fusion protein of single-chain variable domain fragments for treatment of myasthenia gravis

Fangfang Li, Fanping Meng, Quanxin Jin, Changyuan Sun, Yingxin Li, Honghua Li, Songzhu Jin   

  1. Department of Immunology and Pathogenic Biology, College of Medicine, Yanbian University, Yanji, Jilin Province, China
  • Received:2014-03-01 Online:2014-04-25 Published:2014-04-25
  • Contact: Fanping Meng, Ph.D., Department of Immunology and Pathogenic Biology, College of Medicine, Yanbian University, 977# Gongyuan Road, Yanji 133002, Jilin Province, China, fpmeng@ybu.edu.cn.
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 30360100, 30760234, 30860260, 81160373, 81360458.

摘要:

单链可变区片段637是重症肌无力的抗原特异性单链可变区片段。实验将单链可变区片段基因与人血清白蛋白相结合,并在毕赤酵母中表达两者的融合蛋白,以免疫组化染色验证该融合蛋白能与人体肋间肌中神经肌肉接头中的乙酰胆碱受体结合。以竞争性ELISA验证显示融合蛋白对重症肌无力患者血清与乙酰胆碱受体结合的抑制率为2.0-77.4%,且融合蛋白在正常血清中的稳定性约为3d。结果说明单链可变区片段-人血清白蛋白可以作为特异性免疫抑制治疗重症肌无力的可选制剂。

关键词: 神经再生, 重症肌无力, 乙酰胆碱受体, 抗乙酰胆碱受体抗体, 单链Fv, 人血清白蛋白, 融合蛋白, 免疫抑制治疗, 自身免疫性疾病, NSFC grant

Abstract:

Single-chain variable domain fragment (scFv) 637 is an antigen-specific scFv of myasthenia gravis. In this study, scFv and human serum albumin genes were conjugated and the fusion protein was expressed in Pichia pastoris. The affinity of scFv-human serum albumin fusion protein to bind to acetylcholine receptor at the neuromuscular junction of human intercostal muscles was detected by immunofluorescence staining. The ability of the fusion protein to block myasthenia gravis patient sera binding to acetylcholine receptors and its stability in healthy serum were measured by competitive ELISA. The results showed that the inhibition rate was 2.0–77.4%, and the stability of fusion protein in static healthy sera was about 3 days. This approach suggests the scFv-human serum albumin is a potential candidate for specific immunosuppressive therapy of myasthenia gravis.

Key words: nerve regeneration, myasthenia gravis, acetylcholine receptor, anti-acetylcholine receptor antibody, single-chain variable domain fragment, human serum albumin, fusion protein, immunosuppressive therapy, autoimmune disease, NSFC grant, neural regeneration