烟曲霉Asp f3和Asp f4的表达纯化与多克隆抗体的制备  

Expression and Purification of Aspergillus Fumigates Asp f3 and Asp f4 and Preparation of Polyclonal Antibodies Against Asp f3 and Asp f4

作　　者：高岩[1,2] 霍克克[1] 季朝能[1,2] 唐海平[3] 徐万祥[3] 谢毅[1,2] 顾少华[1,2] 

机构地区：[1]复旦大学生命科学学院,遗传工程国家重点实验室,上海200438 [2]上海市工业菌株工程技术研究中心,上海200438 [3]上海市计划生育科学研究所,国家人口和计划生育委员会重点实验室,上海200032

出　　处：《复旦学报（自然科学版）》2015年第5期601-608,共8页Journal of Fudan University：Natural Science

基　　金：国家重点基础研究发展计划(2013CB531603);上海市科学技术委员会项目(13DZ2252000)

摘　　要：侵袭性曲霉病以烟曲霉在肺部定植、侵袭为主,具有较高的致死率.治疗侵袭性曲霉病的关键是早期确诊.目前临床上用于诊断侵袭性曲霉病的血清学方法是检测血清中半乳甘露聚糖循环抗原的GM试验,在敏感性和特异性方面存在局限性.为了用抗重组蛋白多克隆抗体鉴定和分析烟曲霉Asp f3和Asp f4特异性和曲霉广谱性表位,构建检测可疑血清样本中相应抗体的表位嵌合肽,以诊断侵袭性曲霉病.本研究表达纯化了烟曲霉Asp f3和Asp f4重组蛋白,制备了IgG抗体滴度均在1.28×106以上的高滴度的兔抗重组蛋白多克隆抗体.Invasive Aspergillosis（IA）has high mortality forcolonization and invasionin lungsof Aspergillusfumigatu,early diagnosis is critical for treatment of IA.Serological method currently used in the clinical diagnosis of invasive aspergillosis is GM test：a detection of serum galactomannan circulating antigen,which is limited in terms of sensitivity and specificity.For the purpose of identifying and analyzing the specific epitopes of Asp f3 and Asp f4,finding out the broad-spectrum epitopes of Aspergillus and developing the chimeric antigen peptides for IA diagnosis,we expressed and purified Aspergillus fumigates recombinant protein Asp f3 and Asp f4,and then prepared high titer（higher than 1.28×105）polyclonal rabbit antibodies against the two recombinant protein respectively in this study.

关 键 词：烟曲霉 ASP F3 ASP f4 表达纯化 抗体制备 

分 类 号：Q352[生物学—遗传学]