鲍曼不动杆菌外膜蛋白W多克隆抗体的制备及鉴定

PREPARATION AND IDENTIFICATION OF ANTI-ACINETOBACTER BAUMANNII OUTER MEMBRANE PROTEIN W POLYCLONAL ANTIBODY

目的:制备鲍曼不动杆菌外膜蛋白W(OmpW)多克隆抗体,为进一步研究OmpW在鲍曼不动杆菌多重耐药及致病机制中的作用提供基础。方法:在Pubmed检索OmpW信息,选取外膜蛋白W特定的氨基酸区域合成多肽后偶联蓝色载体,并作为免疫原免疫新西兰大白兔,得到抗血清并进行纯化。采用5株鲍曼不动杆菌临床分离株的菌体蛋白对抗体进行鉴定。结果:偶联多肽的蓝色载体构建成功,经动物免疫后得到抗血清,经纯化后得到高纯度的OmpW多克隆抗体。经临床分离鲍曼不动杆菌菌体蛋白验证,证实得到理想的兔抗OmpW多克隆抗体。结论:通过多肽合成制备免疫原方法及动物免疫,成功获得了特异性高的多克隆抗体,为进一步研究鲍曼不动杆菌OmpW蛋白在不同鲍曼不动杆菌临床株中的分布特征及其抗体的保护效应奠定了基础。

Objective： To prepare Acinetobacter baumannii outer membrane protein W（ OmpW）polyclonal antibody,and further provide basis for exploring the effects of Omp W in the pathogenesis of Acinetobacter baumannii. Methods： The gene and protein information of OmpW were retrieved in Pub Med. Amino acid region of outer membrane protein W was selected to synthesize polypeptide which was then coupled with blue vector. The synthesized protein was used as immunogen to immune New Zealand white rabbits and the anti-sera was obtained,purified and identified using mycoprotein abstracted from five clinical acinetobacter baumannii isolates. Results： The blue vector coupled with synthesized polypeptide vector was constructed successfully. After immunization of animals,anti-sera was obtained successfully. The purified antibody was verified by mycoprotein abstracted fromclinical Acinetobacter baumannii isolates,which confirmed that the ideal anti-OmpW antibody was prepared successfully. Conclusion： The ideal OmpW polyclonal antibody was generated using polypeptide synthesization and animal immunization method,which provide a basis for further investigating the distribution features of Omp W among different clinical isolates and the protective effects of its polyclonal antibody.