CAJ | 학술논문

目的:对A族链球菌Fba蛋白McAb2所对应的表位进行定位.方法:以初步定位的表位区段为线索合成三段重叠多肽,dot-ELISA检测McAb2与合成肽的亲合力,并以McAb2为靶分子,利用噬菌体随机七肽库进行亲和筛选,用竞争ELISA鉴定阳性克隆.结果:dot-ELISA检测结果表明Fba_(100-112)肽段与McAb2的结合能力最强.经过3轮亲和筛选后,随机挑选20个噬菌体克隆,竞争ELISA对其与McAb2的亲和力做检测,其中12个克隆显示较强的阳性结果.阳性克隆DNA测序,与Fba基因第100位氨基酸至110位氨基酸中ITPDL同源性较高.结论:通过dot-ELISA将A族链球菌Fba蛋白McAb2对应的表位定位于100～112位氨基酸,结果同噬菌体随机肽库筛选的核心氨基酸位置吻合.为进一步研制表位肽疫苗、研究Fba蛋白及其单克隆抗体的生物学功能奠定了基础.
목적:대A족련구균Fba단백McAb2소대응적표위진행정위.방법:이초보정위적표위구단위선색합성삼단중첩다태,dot-ELISA검측McAb2여합성태적친합력,병이McAb2위파분자,이용서균체수궤칠태고진행친화사선,용경쟁ELISA감정양성극륭.결과:dot-ELISA검측결과표명Fba_(100-112)태단여McAb2적결합능력최강.경과3륜친화사선후,수궤도선20개서균체극륭,경쟁ELISA대기여McAb2적친화력주검측,기중12개극륭현시교강적양성결과.양성극륭DNA측서,여Fba기인제100위안기산지110위안기산중ITPDL동원성교고.결론:통과dot-ELISA장A족련구균Fba단백McAb2대응적표위정위우100～112위안기산,결과동서균체수궤태고사선적핵심안기산위치문합.위진일보연제표위태역묘、연구Fba단백급기단극륭항체적생물학공능전정료기출.
Objective:To identify monoclonal antibody McAb2 recognizing epitope of Fba of GAS.Methods:The overlapped peptides were synthesized and their abilities to bind McAb2 were detected by dot-ELISA.The predominance amino acids specific for McAb2 were screened using phage 7 peptide library.Results:The result by dot-ELISA analysis demonstrated that the synthetized peptide,amino-acid residues 100-112~(th),could bind McAb2 with high affinity.The predominance amino acids specific for McAb2 were ITPDL,which was located in 100-110~(th)aa of Fba by panning with phage 7 peptide library.Conclusion:The domain and the predominance amino acids of Fba recognized by McAb2 is determined.The results would contribute to the research of the role of Fba on the pathogenic mechanism of GAS,the identification of function of McAb2,and the development of epitope-peptide vaccine.