摘要
利用基因工程技术在大肠杆菌BL21(DE3)中表达白喉毒素无毒突变体CRM_(197),经变性条件下亲和纯化后,作为蛋白载体与流感抗原M2e经BMPH偶联,制备CRM_(197)-M2e结合物,用其免疫BALB/c小鼠,采用间接ELISA法测定血清中M2e特异性IgG抗体.结果表明,重组CRM_(197)在大肠杆菌中成功表达,优化后蛋白表达量约为250±30 mg/L,纯度达95%以上;所制CRM_(197)-M2e结合物可有效免疫BALB/c小鼠,其刺激产生的M2e特异性IgG抗体滴度分别是健康组、M2e免疫对照组、CRM_(197)免疫对照组及CRM_(197)+M2e混合组的430.5,32,181和215倍.
Express the nontoxic mutant of diphtheria toxin（CRM（197）） in E.coli BL21（DE3） by gene engineering technique and purify the protein by affinity chromatography under denaturation.As the carrier protein,recombinant CRM（197） was conjugated with influenza antigen M2e via BMPH to prepare the conjugate vaccine to immune BALB/c mice.Determine the M2e specific IgG in sera by indirect ELISA.The results showed that r CRM_（197） was expressed in E.coli system successfully with optimized expression level（250±30 mg/L） and high purity（95%）.The conjugate vaccine CRM（197）-M2e prepared by chemical coupling in denaturation could effectively immunize BALB/c mice.And the M2e-specific IgG antibody titer elicited by CRM_（197）-M2e conjugate vaccine was significantly higher than that in healthy,M2e,CRM_（197） and M2e＋CRM_（197） mixed groups by 430.5,32,181 and 215 times.
出处
《过程工程学报》
CAS
CSCD
北大核心
2017年第5期1054-1058,共5页
The Chinese Journal of Process Engineering
基金
教育部博士点基金资助项目(编号:120120181110036)
