Yiping Xun1, Peng Chen1, Hai Yan1, Weikang Yang1, Lili Shi1, Guangyu Chen2, Hongwu Du3. 1. School of Chemistry and Biotechnology Engineering, University of Science & Technology Beijing, Beijing 100083, China. 2. ImmunoHunt Corporation, 139 Fengtai Rd, Beijing 100071, China. 3. School of Chemistry and Biotechnology Engineering, University of Science & Technology Beijing, Beijing 100083, China. Electronic address: hongwudu@ustb.edu.cn.
Abstract
OBJECTIVE: This study is intended to screen potential antigen for Behcet's disease (BD) by using human microvascular endothelial cells (HUVEC). METHODS: Following cell-based indirect immunofluorescence assay with sera from BD patients, proteins extracted from HUVEC were separated and detected by Western blotting. Then the target protein was identified by LC-MALDI-TOF/TOF, the recombinant target protein was expressed, purified and then used as coating antigen to test the prevalence of autoantibodies in patient's sera. RESULTS: The Western blotting result showed that some patients' sera could react with a protein band with about 30 kDa of molecular weight, which was further identified as prohibitin by mass spectrometry. The prevalence of serum antibodies against recombinant human prohibitin was detected in 16 of 58 BD patients (28%) but none in healthy controls.
OBJECTIVE: This study is intended to screen potential antigen for Behcet's disease (BD) by using human microvascular endothelial cells (HUVEC). METHODS: Following cell-based indirect immunofluorescence assay with sera from BDpatients, proteins extracted from HUVEC were separated and detected by Western blotting. Then the target protein was identified by LC-MALDI-TOF/TOF, the recombinant target protein was expressed, purified and then used as coating antigen to test the prevalence of autoantibodies in patient's sera. RESULTS: The Western blotting result showed that some patients' sera could react with a protein band with about 30 kDa of molecular weight, which was further identified as prohibitin by mass spectrometry. The prevalence of serum antibodies against recombinant humanprohibitin was detected in 16 of 58 BDpatients (28%) but none in healthy controls.