CAJ | 학술논문

目的从全人源抗鼻咽癌噬菌体抗体库中筛选特异性单链抗体(ScFv),并对其特异性进行鉴定.方法通过噬菌体表面展示技术把ScFv表达在噬菌体表面,以鼻咽癌细胞作为抗原,用抗原递减法,通过"吸附-洗脱-扩增"过程筛选并富集特异性抗体,及ELISA筛选,获得特异阳性克隆进行免疫组化鉴定并测序.结果通过对抗体库进行三轮正负淘洗和富集后,随机挑选4212个克隆进行ELISA,发现3个克隆对CNE2呈强阳性反应,而与人正常细胞系HUVEC等呈弱阳性反应或不反应.对克隆HNSAO33进一步进行免疫细胞化学验证,结果与ELISA反应一致;免疫组织化学鉴定表明克隆HNSAO33与鼻咽癌组织和鼻咽组织阳性率的差别有统计学意义.结论通过淘选富集、ELISA和免疫化学鉴定获得特异性较强的噬菌体克隆,为鼻咽癌发病机制的研究和临床诊断以及治疗奠定了基础.
목적 종전인원항비인암서균체항체고중사선특이성단련항체(ScFv),병대기특이성진행감정.방법 통과서균체표면전시기술파ScFv표체재서균체표면,이비인암세포작위항원,용항원체감법,통과"흡부-세탈-확증"과정사선병부집특이성항체,급ELISA사선,획득특이양성극륭진행면역조화감정병측서.결과 통과대항체고진행삼륜정부도세화부집후,수궤도선4212개극륭진행ELISA,발현3개극륭대CNE2정강양성반응,이여인정상세포계HUVEC등정약양성반응혹불반응.대극륭HNSAO33진일보진행면역세포화학험증,결과여ELISA반응일치;면역조직화학감정표명극륭HNSAO33여비인암조직화비인조직양성솔적차별유통계학의의.결론 통과도선부집、ELISA화면역화학감정획득특이성교강적서균체극륭,위비인암발병궤제적연구화림상진단이급치료전정료기출.
Objective To screen the anti-nasopharyngeal carcinoma scFv from a human anti-nasopharyngeal carcinoma single-chain phage antibody library, and identify its characteristics. Methods The single-chain phage antibody library was subjected to three rounds of positive and negative cell panning and enrichment, and then it was selected by ELISA. The binding specificity of phage antibodies with naso-pharyngeal carcinoma cells was confirmed by immunohistochemistry. Results After panning, enrichment and testing by ELISA, 3 phage an-tibody clones reacting with CNE2 more strongly than HUVEC and NP69 were picked out from 4212 clones. One clone, HNSAO33, was fur-ther analyzed after DNA sequencing. The results of immunohistochemistry with cultured cells were similar to those of ELISA. HNSAO33 spe-cifically reacted to nasopharyngeal carcinoma cells in most human nasopharyngeal carcinoma tissue sections except a few human normal naso-pharyngeal tissue sections. The distinction of positive rates was of a great statistical significance. Conclusion ELISA and immunohisto-chemistry results confirmed HNSAO33 specifically bind with nasopharyngeal carcinoma cells. The seFv fragment against nasopharyngeal carci-noma may be further developed and applied in clinical diagnosis and therapy of nasopharyngeal carcinoma.