CAJ | 학술논문

目的用噬菌体展示肽库技术筛选甲型肝炎(甲肝)病毒抗原模拟表位,为病毒抗原决定簇定位探索可行方法.方法用纯化的抗甲肝病毒单克隆抗体,对噬菌体展示12肽库进行3轮“吸附-洗脱-扩增”筛选,随机挑取10个克隆,用酶联免疫吸附法(ELISA)对噬菌体克隆进行抗原性鉴定、竞争抑制鉴定及DNA序列测定分析,推导出展示肽氨基酸序列并与甲肝病毒(HAV)代表株结构蛋白氨基酸序列比较.结果 10个噬菌体克隆ELISA检测全为阳性,9个具有一致序列,与HAVHM175株结构蛋白中和活性表位之一:VP1 157-171区具有类似序列,另一株噬菌体克隆在HAVHM175中未发现类似序列,结果表明这些展示肽可能是HAV抗原模拟表位.结论用噬菌体展示肽库技术筛选得到了HAV模拟表位,为开展病毒模拟表位研究打下了基础.
목적 용서균체전시태고기술사선갑형간염(갑간)병독항원모의표위,위병독항원결정족정위탐색가행방법.방법 용순화적항갑간병독단극륭항체,대서균체전시12태고진행3륜“흡부-세탈-확증”사선,수궤도취10개극륭,용매련면역흡부법(ELISA)대서균체극륭진행항원성감정、경쟁억제감정급DNA서렬측정분석,추도출전시태안기산서렬병여갑간병독(HAV)대표주결구단백안기산서렬비교.결과 10개서균체극륭ELISA검측전위양성,9개구유일치서렬,여HAVHM175주결구단백중화활성표위지일:VP1 157-171구구유유사서렬,령일주서균체극륭재HAVHM175중미발현유사서렬,결과표명저사전시태가능시HAV항원모의표위.결론 용서균체전시태고기술사선득도료HAV모의표위,위개전병독모의표위연구타하료기출.
Objective A 12 mer phage display peptide library was used to identify hepatitis A virus mimotopes of antigenic determinants,to provide the feasibility of virus epitope mapping by using this approach.Methods Using purified anti-hepatitis A virus monoclonal antibody as affinity selective molecule,phage display peptide library was biopanned and positive clones were selected by ELISA,competition assay and DNA sequencing.Results 10 ELISA positive clones were chosen for DNA sequencing,and the displayed peptide sequences were deduced.9 of them showed identical nucleotide sequence,and similarity in their amino acid sequence with VP1 of HAV HM175 was found,but no sequence homology was found between the other phage clone and the capsid proteins of HAV.Those peptides may behave as mimotopes of HAV.Conclusion The mimotope of HAV was selected by using phage display peptide library screening.The results provide the potential of this method to search for the mimotopes of the virus.