中华皮肤科杂志 ›› 2008, Vol. 41 ›› Issue (11): 742-744.

• 论著 • 上一篇    下一篇

大疱性类天疱疮患者血清抗BP180分子不同表位自身抗体定量分析及亲和力分析

王刚 刘玉峰   

  1. 第四军医大学西京医院
  • 收稿日期:2007-09-24 修回日期:2007-10-20 发布日期:2008-11-15
  • 通讯作者: 王刚 E-mail:xjwgang@fmmu.edu.cn

Quantification and affinity assessment of autoantibodies against different epitopes of BP180 from patients with bullous pemphigoid

  

  • Received:2007-09-24 Revised:2007-10-20 Published:2008-11-15

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摘要： 目的 探讨大疱性类天疱疮（BP）患者血清抗BP180分子不同表位的自身抗体量。方法 制备BP180分子NC16A片段的不同抗原表位区NC16A-1、NC16A-2和NC16A-3，利用柱亲和层析的方法从10例BP患者血清中分别纯化抗不同表位区的自身抗体，定量，并用硫氰酸盐洗脱法测定抗NC16A-1、抗NC16A-2和抗NC16A-3自身抗体的相对亲和力。结果 经亲和层析纯化获得针对BP180-NC16A不同表位区的自身抗体，从20 mg总IgG中纯化抗NC16A-1、NC16A-2和NC16A-3自身抗体的产量分别为（49.0 ± 20.7） μg、（117.7 ± 22.4） μg和（39.5 ± 18.9） μg。抗NC16A-2自身抗体的量和亲和力水平均明显高于其他两个表位区的自身抗体。结论 BP致病性自身抗体所识别的主要抗原表位可能位于BP180分子的NC16A-2表位区（aa507-aa520）。

关键词: 类天疱疮, 大疱性;自身抗体;抗原;表位;抗体亲和力

Abstract: Objective To purify and quantify the autoantibodies against various epitopes within BP180-NC16A domain from the sera of patients with bullous pemphigoid （BP）. Methods Three epitopes within BP180-NC16A domain, NC16A-1, NC16A-2 and NC16A-3, were prepared. Blood samples were obtained from 10 patients who were diagnosed as active BP by clinical, pathological and immunofluorescence examination. Autoantibodies against these epitopes were purified with affinity chromatography column from the sera of these patients with active BP, and quantified separately. The relative binding affinity of autoantibodies to NC16A-1, NC16A-2 and NC16A-3 was measured using thiocyanate elution method. Results The autoantibodies against NC16A-1, NC16A-2 and NC16A-3 were successfully purified from the sera of patients. On average, the amount was 49.0 ± 20.7 μg, 117.7 ± 22.4 μg and 39.5 ± 18.9 μg respectively for autoantibodies to NC16A-1, NC16A-2 and NC16A-3 purified from a portion of serum containing about 20 mg IgG. Both the amount and binding affinity of anti-NC16A-2 autoantibody were significantly higher than those of anti-NC16A-1 and anti-NC16A-3 autoantibodies. Conclusion BP180 NC16A-2 （aa507-aa520） may be the major epitope recognized by pathogenic autoantibodies in patients with BP.

Key words: Bullous pemphigoid, Autoantibody