日本血吸虫未成熟卵单链抗体库的构建、筛选及初步应用  

Construction,Screening and Application of Single-chain Fv Antibody Library Against Immature Egg of Schistosoma japonicum

作　　者：何卓[1] 汪世平[1] 肖小芹[1] 曾少华[2] 刘明社[3] 李林[1] 周帅锋[1] 

机构地区：[1]中南大学湘雅医学院寄生虫学系,长沙410078 [2]湖南师范大学第三附属医院,长沙410006 [3]山西省长治医学院寄生虫学教研室,长治046000

出　　处：《生物化学与生物物理进展》2008年第8期921-928,共8页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展计划(973)(2007CB513108);国家高技术研究发展计划(863)(2004AA2Z3530);湖南省"十一.五"重大专项(2006SK1001);湖南省"十一.五"重点学科建设专项经费(07-985-2)联合资助项目~~

摘　　要：运用噬菌体展示技术构建日本血吸虫未成熟虫卵可溶性抗原(SIEA)单链抗体(scFv)表达文库,以天然分子候选疫苗SIEA26～28ku为靶抗原筛选SIEA单链抗体库,获得特异性单链抗体.并将该scFv基因亚克隆至原核高效表达载体PET32a,诱导SIEA26～28ku特异性scFv大量表达.随后以此为探针筛选日本血吸虫尾蚴cDNA文库,以期获得SIEA26～28ku天然分子候选疫苗相关的编码基因.结果显示,所获得的SIEA26～28ku特异性scFv,表达量高,采用该探针初步筛选出相关基因核糖体蛋白S4.SIEA26～28ku特异性scFv的获得,为进一步筛选、分析鉴定抗日本血吸虫病天然分子候选疫苗SIEA26～28ku的编码基因奠定了基础.Single-chain Fv （scFv） antibody library against SIEA （soluble immature egg antigen） of Schistosoma japonicum was constructed by phage display technology. Specific SIEA26-28 ku scFv was obtained by screening the SIEA scFv library using the natural molecular candidate vaccine, SIEA26-28 ku. The specific scFv fragment was subcloned to prokaryotic expression vector PET32a and induced the expression of soluble specific scFv in high level. Subsequently, specific scFv as a probe was used to screen the cercaria eDNA library of Schistosoma japonicum to get the coding genes ofSIEA26-28 ku molecules. Results showed that specific SIEA26-28 ku scFv with high level expression was achieved. The corresponding gene, ribosomal protein S4 was obtained by initially screening the cercaria eDNA library with the specific SIEA26-28 ku scFv. The obtaining of specific SIEA26-28 ku scFv lays the foundation for further screening and identification of the coding genes of SIEA26-28 ku, the natural molecular candidate vaccine against schistosomiasis.

关 键 词：日本血吸虫 未成熟虫卵可溶性抗原(SIEA) 单链抗体(scFv) 噬菌体展示抗体库 CDNA文库 

分 类 号：R383[医药卫生—医学寄生虫学]