大肠杆菌表达的破伤风毒素C片段的免疫原性分析  

Immunogenicity of Tetanus Toxin Fragment C Expressed in E.coli

作　　者：谭亚军[1] 雷殿良[1] 张庶民[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《中国生物制品学杂志》2004年第5期261-263,共3页Chinese Journal of Biologicals

摘　　要：目的　分析大肠杆菌表达的破伤风毒素C片段 (简称TTC)的免疫原性。方法　利用分子生物学技术在大肠杆菌中表达破伤风毒素C片段 ,用阴离子交换层析和金属离子螯合亲和层析方法进行蛋白纯化。参照2 0 0 0年版《中国生物制品规程》的方法检测表达产物的效价。用间接血凝实验检测免疫动物血清中的破伤风抗体单位。结果　重组蛋白的表达量占菌体总蛋白的 15 %。纯化后蛋白纯度达到 96 5 %。免疫效价为 18 70 6IU mg ,ED50 为 2 0 μg。 1μgTTC经 4次免疫能诱导机体产生足够的抗体 ,保护动物免受破伤风毒素的攻击。 结论　重组蛋白具有良好的免疫原性 。Objective To analyze the immunogenicity of tetanus toxin fragment C expressed in E.coli.Methods Express tetanus toxin fragment C in E.coli by molecular biological technique and purify the expressed product by anion exchange and Ni^(2+)-chelate affinity chromatography. Determine the potency of expressed product by the method in Chinese Requirements for Biologics (2000). Detect the tetanus antiserum units of immunized animals by indirect hemagglutination assay.Results The expressed product contained 15% of total somatic protein and reached a purity of 96.5% after purification. The immune potency and ED_(50) of the expressed product were 18.706 IU/mg and 20 μg respectively. The immunization with 4 doses (1 μg for each) of the expressed tetanus toxin fragment C induced sufficient antibody to protect animals from challenge with tetanus toxin.Conclusion The fragment C of tetanus toxin showed good immunogenicity. The study laid a foundation of development of recombinant tetanus vaccine.

关 键 词：大肠杆菌 表达 破伤风毒素 免疫原性 抗体 

分 类 号：R517.3[医药卫生—内科学] R392-33[医药卫生—临床医学]