应用Vero细胞法测定重组减毒绿脓杆菌外毒素A的残余毒力  

Determinate the toxicity of attenuated rETA by using Vero cell assay

作　　者：马霄[1] 田霖[1] 张庶民[1] 雷殿良[1] 

机构地区：[1]中国药品生物制品检定所血清室,北京100050

出　　处：《微生物学免疫学进展》2003年第4期47-49,共3页Progress In Microbiology and Immunology

摘　　要：绿脓杆菌外毒素A(ETA)有ADP -核糖基转移酶的活性 ,在真核细胞中通过催化ADP -核糖基团从NAD+转移到EF 2 (延长因子 2 ) ,使蛋白质合成终止 ,致细胞死亡 ,其作用机制、机理与白喉毒素相同。本文应用Vero细胞法测定了绿脓杆菌外毒素A的毒性 ,发现Vero细胞对绿脓杆菌外毒素A的敏感度高达 3.0× 10 -5mg/ml,且精密度高 ,准确性好 ,是一种有效、可行的绿脓杆菌外毒素毒力的测定方法。对 3批经基因工程减毒后样品的残余毒力进行了测定 ,发现样品基本无细胞毒性 ,毒力比原毒素低 2× 10In eukaryotic cells, Pseudomonas aeruginosa exotoxin A (ETA) catalyses the transfer of the ADP ribose moiety from NAD + to elongation factor 2 (EF 2). As aresult, protein synthesis ceased, and intoxicated cells died since elongation of polypeptide chains no longer occursed.Wilkens K etc. Revealed that diphteria toxin also had ADP ribosyltransferase activity, and proved that they have the same mechanism.So, we used Vero cell method to assay the toxicity of ETA. The sensitivity of Vero cell to ETA was about 3.0×10 -5 mg/ml and the specificity and precision of Vero cell method was good. Then, we used this method to test the toxicity of the recombinant attenuated ETA for three lots of preparation and found that the samples had almost no cell toxicity, which were 2×10 15 times lower than that of standard ETA.

关 键 词：绿脓杆菌外毒素A ETA VERO细胞 毒力测定 重组 残余毒力 

分 类 号：R96[医药卫生—药理学]