CAJ | 학술논문

目的利用噬菌体随机七肽库技术筛选人膀胱癌相关表面标志物Her2、Survivin表位模拟肽.方法分别以Her2、Survivin多克隆抗体为固相筛选分子,对噬菌体随机七肽库进行3轮生物淘洗,随机挑选单克隆噬菌体进行酶联免疫吸附试验(ELISA)分析检测其结合力,竞争抑制实验鉴定其阳性克隆,对结合力较好的噬菌体提取DNA并测序,寻找比较保守的核心序列,合成多肽,最后鉴定合成多肽的结合力.结果经噬菌体随机七肽库3轮淘洗后,特异性噬菌体模拟肽得到富集,分别挑选Her2、Survivin20个克隆,经DNA测序分析,Her2较保守的序列为ACT ACG GCT GAG AAT AAT GAG,Survivin为TCT CCT CCT CAT CTT TCG CAG,合成多肽,Her2为Thr-Thr-Ala-Glu-Asn-Asn-Glu(TTAENNE),Survivin为Ser-Pro-Pro-His-Leu-Ser-Gln(SPPHLSQ),ELISA分析具有良好结合力.结论利用噬菌体随机七肽库成功筛选到人膀胱癌相关表面标志物Her2、Survivin表位模拟肽,从而为今后膀胱癌多肽疫苗的研制提供良好的基础和条件.
목적 이용서균체수궤칠태고기술사선인방광암상관표면표지물Her2、Survivin표위모의태.방법 분별이Her2、Survivin다극륭항체위고상사선분자,대서균체수궤칠태고진행3륜생물도세,수궤도선단극륭서균체진행매련면역흡부시험(ELISA)분석검측기결합력,경쟁억제실험감정기양성극륭,대결합력교호적서균체제취DNA병측서,심조비교보수적핵심서렬,합성다태,최후감정합성다태적결합력.결과경서균체수궤칠태고3륜도세후,특이성서균체모의태득도부집,분별도선Her2、Survivin20개극륭,경DNA측서분석,Her2교보수적서렬위ACT ACG GCT GAG AAT AAT GAG,Survivin위TCT CCT CCT CAT CTT TCG CAG,합성다태,Her2위Thr-Thr-Ala-Glu-Asn-Asn-Glu(TTAENNE),Survivin위Ser-Pro-Pro-His-Leu-Ser-Gln(SPPHLSQ),ELISA분석구유량호결합력.결론 이용서균체수궤칠태고성공사선도인방광암상관표면표지물Her2、Survivin표위모의태,종이위금후방광암다태역묘적연제제공량호적기출화조건.
Objective To screen Her2 and Survivin mimic peptides that were associated with bladder cancer from Ph.D.-7 phage display peptide library.Methods Ph.D.-7 phage display peptide library was biopanned for 3 rounds by using Her2 and Survivin polyclonal antibody and then phage titer was detected.Enzyme linked immunosorbent assay (ELISA) dection and competitive supression test were performed to detect the binding force of random-selected monoclonal phages.DNA extracted from phages with better binding activity were sequenced and got the basic group of core sequence that was appeared more often relatively.Then the polypeptide was synthesized and ELISA detection and competitive supression test were performed again to detect the binding force.Results After 3 rounds of biopanning,the titer of phages were increased gradually.Select 20 monoclonal plaque from titering plates of Her2 and Survivin respectively.After the sequence of DNA,the core sequence of her2 was ACT ACG GCT GAG AAT AAT GAG and the sequence of Survivin was TCT CCT CCT CAT CTT TCG CAG.Then polypeptides were synthesized successfully.The sequence of Her2 polypeptide was Thr-Thr-Ala-Glu-Asn-Asn-Glu (TTAENNE) and the sequence of Survivin polypeptide was Ser-Pro-Pro-His-Leu-Ser-Gln(SPPHLSQ).Conclusion Ph.D.-7 phage display peptide library was used to screen Her2 and Survivin mimic peptides successfully,which would supply a good basement and condition for the advanced reseach about polypeptide vaccinum of bladder cancer.