中国免疫学杂志2018，Vol.34Issue(2)：233-238,6.DOI:10.3969/j.issn.1000-484X.2018.02.015
抗铜绿假单胞菌PcrV蛋白单克隆抗体的筛选和功能验证
Screening and activity verification of monoclonal antibody against PcrV protein of pseudomonas aeruginosa
摘要
Abstract
Objective:To obtain a high specificity and high affinity anti-PcrV protein monoclonal antibody which can be used for the treatment of Pseudomonas aeruginosa infected.Methods: The PcrV gene was amplified by PCR using P.aeruginosa PAO1 genome DNA as the template.The expression vector(pET-28a-PcrV) was constructed and transformed into E.coli BL21(DE3).The re-combinant PcrV protein was expressed by IPTG induction and purified by Ni2+affinity chromatography.The specific binders of PcrV were screened by phage display.The genes encoding VH and VL were amplified respectively by PCR using the plasmid of positive clone as the template.Then the recombinant expression vectors were constructed and transfected into 293E cells.Monoclonal antibody were purified by the Protein A affinity resin from the culture supernatants.The affinity of antibody was detected by ELISA and the function of YG5 was verified in murine pneumonia model caused by P.aeruginosa.Results: Recombinant PcrV protein was expressed and purified.A full human monoclonal antibody(named as YG5) against PcrV was obtained by phage display.The results of ELISA showed that YG5 had a high affinity with EC50=61 ng/ml.Furthermore,it was found that YG5 could protect mice from infection caused by P.aeruginosa.Conclusion:Our findings present a novel human monoclonal antibody YG5 against PcrV,which inhibits the infection casued by P.aeruginosa and may be a potential drug for treatment of P.aeruginosa infection.关键词
铜绿假单胞菌/PcrV蛋白/单克隆抗体/急性肺炎模型Key words
Pseudomonas aeruginosa/Protein PcrV/Monoclonal antibody/Acute pneumonia model分类
医药卫生引用本文复制引用
管章春,刘方杰,刘成华,高亚萍,沈倍奋,杨光..抗铜绿假单胞菌PcrV蛋白单克隆抗体的筛选和功能验证[J].中国免疫学杂志,2018,34(2):233-238,6.基金项目
本文受国家自然科学基金资助(31170074). （31170074）
