目的:筛选CD20分子的模拟表位肽,构建针对CD20分子的治疗性疫苗,以期为淋巴瘤以及其它B细胞相关性疾病的治疗提供新的方向.方法:利用噬菌体随机呈现肽库筛选技术,以人淋巴细胞分化抗原CD20 mAb Rituximab为靶点,筛选CD20分子的模拟表位肽.通过ELISA方法检测筛选出的阳性噬菌体与Rituximab的特异性结合,并以竞争性结合实验检测筛选出的阳性噬菌体与Raji细胞表面的CD20分子竞争结合Rituximab的能力.最后以Sanger双脱氧链终止法测定DNA序列,推断其氨基酸序列.结果:成功筛选出针对CD20 mAb Rituximab的阳性噬菌体,获得了CD20分子的模拟表位肽QDKLTQWPKWLE.获得的阳性噬菌体能够与Rituximab特异性结合,并且表达该表位的噬菌体可以竞争性抑制Rituximab与天然CD20分子的结合.结论:CD20分子的抗原表位肽QDKLTQWPKWLE能够与mAb Rituximab特异性结合,与天然CD20分子竞争性结合mAb Rituximab,并具有潜在的应用价值.
Screen of CD20 mimotope using monoclonal antibody Rituximab
AIM: To seek a new strategy for the treatment of B-cell lymphomas by screening the mimotope of CD20 and constructing a novel treatment vaccine against CD20. METHODS: We screened the CD20 mimotope using CD20 monoclonal antibody Rituximab as the target. The special bind of the phage to Rituximab was detected by ELISA. To characterize the competitive binding ability between the screened positive phage and CD20 to Rituximab,competitive ELISA was used. DNA sequence was analyzed by Sanger method. RESULTS: The positive phage binding to the CD20 monoclonal antibody Rituximab was successfully screened and the CD20 mimotope QDKLTQWPKWLE was obtained. The positive phage bound specifically to Rituximab. Moreover,the positive phage inhibited the binding of Rituximab to CD20. CONCLUSION: The CD20 mimotope QDKLTQWPKWLE binds specifically to Rituximab and it competes to bind Rituximab with CD20.
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