重组表达A型肉毒毒素重链C端多肽及其在多抗血清中的抗原性研究

Recombinant expression of botulinum toxin type A heavy chain C fragment and its antigenicity in polyclonal antiserum

目的对A型肉毒毒素重链C端多肽原始基因序列进行密码子优化后转入大肠杆菌系统进行原核表达并纯化,研究其在多抗血清中的抗原性。方法使用http://www.jcat.de/数据库等生物信息学技术对A型肉毒毒素重链C端多肽原始基因序列进行密码子优化后装入原核表达载体p ET-22b(+)中,转入大肠杆菌工程菌株BL21(DE3)表达并纯化,通过动物实验研究其在多抗血清中的抗原性。结果成功表达并纯化了A型肉毒毒素重链C端多肽,并对其抗原性进行验证。结论 A型肉毒毒素重链C端多肽作为抗肉毒毒素的二代亚单位疫苗组分以及肉毒类毒素的替代品,在制备以及激起中和抗体的方面具有明显的优势。

Botulinum toxin A（BONT/A） and its related bioproducts have been widely used in many medicaltreatment and have got remarkable achievement. However, the counterterrorism concerns of Botulism of type A aremounted every day. Here, we aimed to express botulinum toxin type A heavy chain C fragment and evaluate itsantigenicity in polyclonal antiserum. The Botulinum neurotoxin type A heavy chain C fragment gene codon wasoptimized and then expressed in E. coli. The antigenicity of the recombinant heavy chain C fragment（r AHc） wasinvestigated by animal experiment. The result showed that r AHc has a dramatic advantage in take place ofBotulinum toxoid A for the preparation of polyclonal antiserum and subunit vaccine of Botulinum toxin A.