CAJ | 학술논문

目的：建立酶联免疫吸附分析方法（ ELISA ）检测待测样品中的人源化抗体MIL50。方法用原核表达的重组蓖麻毒素（ ricin） A链（ RTA）突变体（ RiVax）为包被抗原，与辣根过氧化物酶（ HRP）标记的羊抗人IgG建立的夹心ELISA，检测生物样品中的人源化抗体MIL50。结果以蓖麻毒素和RiVax包被抗原，ELISA检测MIL50的结果显示，RiVax可与MIL50特异性结合，结合能力不低于全毒素蛋白，可以用于MIL50的定量分析。 RiVax的包被浓度为3 mg/L，对于溶解于含有吐温20（Tween 20）磷酸缓冲液（PBST）中的MIL50的检测灵敏度可达0．030 mg/L；与PBST样品相比较，含有同样浓度MIL50的大鼠血清ELISA的阳性结果明显减弱，不同的大鼠组织匀浆对于MIL50的检测有不同程度的干扰。结论夹心ELISA能够有效地用于人源化蓖麻毒素抗体MIL50的检测分析。
목적：건립매련면역흡부분석방법（ ELISA ）검측대측양품중적인원화항체MIL50。방법용원핵표체적중조비마독소（ ricin） A련（ RTA）돌변체（ RiVax）위포피항원，여랄근과양화물매（ HRP）표기적양항인IgG건립적협심ELISA，검측생물양품중적인원화항체MIL50。결과이비마독소화RiVax포피항원，ELISA검측MIL50적결과현시，RiVax가여MIL50특이성결합，결합능력불저우전독소단백，가이용우MIL50적정량분석。 RiVax적포피농도위3 mg/L，대우용해우함유토온20（Tween 20）린산완충액（PBST）중적MIL50적검측령민도가체0．030 mg/L；여PBST양품상비교，함유동양농도MIL50적대서혈청ELISA적양성결과명현감약，불동적대서조직균장대우MIL50적검측유불동정도적간우。결론협심ELISA능구유효지용우인원화비마독소항체MIL50적검측분석。
Objective To develop a sandwich enzyme-linked immunosorbent assay ( ELISA) for the determination of a humanized antibody MIL50.Methods RiVax, a mutant of RTA (a chain of ricin) expressed in E.coli, was used as the coating protein.Horseradish peroxidase (HRP) labeled IgG of goats against humans was used to determine MIL 50 captured by RiVax coated on the plate.Results Compared with ricin, RiVax could bind well with MIL50,indicating it could be used as a coating protein to determine MIL50 instead of holotoxin.Using this method, the detective limit of MIL50 in PBST (PBS containing 0.1%Tween 20) could be as low as 0.030 mg/L.The absorbance value of ELISA for MIL50 in the ser-um and homogenate of the liver , spleen, lung, kidney, muscle of rats was lower than that in PBST .Conclusion The sandwich ELISA is a sensitive method for the analysis of distribution of MIL 50 in rat tissues.