Chen Peng, Yang Weikang, Tian Yaping, Sun Shutao, Chen Guangyu, Zhang ChunYan, Ma Fuxin, Xun Yiping, Shi Lili, Yang Chunhe, Zhao Lanqing, Zhou Yabin, Du Hongwu
School of Chemistry and Biological Engineering, University of Science and Technology Beijing, Beijing, 100083, China.
Department of Clinical Biochemistry, Chinese PLA General Hospital, Beijing, 100853, China.
PLoS One. 2015 Apr 27;10(4):e0124760. doi: 10.1371/journal.pone.0124760. eCollection 2015.
Behçet's disease (BD) is a chronic inflammatory disease with multisystem involvement, and it is listed as a rare disease in the United States but is common in the Middle East, China, and Japan. The aim of this study was to identify novel autoantigens in Chinese patients with BD. First, the candidate autoantigens were screened by Western blotting, and the sequences of putative antigens were identified by LC-MALDI-TOF/TOF mass spectrometry. Next, the screened protein was cloned, expressed and purified. Then, an optimized ELISA was developed, and the serological criteria were evaluated using a large number of confirmed patients. One antigen with a molecular weight of approximately 28 kDa was identified as electron transfer flavoprotein subunit beta (ETFB). Positive reactivity was detected in recombinant human ETFB sera from 38 of 92 BD patients (41 %) and 1 of 90 healthy controls (1 %).
白塞病（BD）是一种累及多系统的慢性炎症性疾病，在美国被列为罕见病，但在中东、中国和日本较为常见。本研究的目的是在中国白塞病患者中鉴定新的自身抗原。首先，通过蛋白质印迹法筛选候选自身抗原，并通过液相色谱-基质辅助激光解吸电离飞行时间/串联飞行时间质谱法鉴定推定抗原的序列。接下来，对筛选出的蛋白质进行克隆、表达和纯化。然后，开发了一种优化的酶联免疫吸附测定（ELISA），并使用大量确诊患者评估血清学标准。一种分子量约为28 kDa的抗原被鉴定为电子传递黄素蛋白β亚基（ETFB）。在92例BD患者中的38例（41%）重组人ETFB血清中检测到阳性反应，而在90例健康对照中的1例（1%）检测到阳性反应。
