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Prokaryotic expression of anti-carcinoembryonic single-chain variable fragment and its value in detection of gastric carcinoma cells

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Author:
No author available
Journal Title:
WORLD CHINESE JOURNAL OF DIGESTOLOGY
Issue:
18
DOI:
10.3969/j.issn.1009-3079.2006.18.006
Key Word:
胃癌细胞;单链抗体;原核表达;癌胚抗原

Abstract: 目的:探讨T84.66单链抗体的原核表达及对6种胃癌细胞系及胃癌组织的特异性亲和力.方法:将抗CEA单链抗体T84.66的cDNA插入噬菌粒载体pCANTAB5E,获得噬菌粒载体T84.66-scFv-pCANTAB5E.将后者转化入E.coli HB2151,经β,D异丙基硫代半乳糖苷(IPTG)诱导表达.采用细胞培养及免疫细胞/组织化学方法,检测胃癌细胞中及石蜡包埋的胃癌组织中的癌胚抗原表达.结果:SDS-PAGE及Western blot证实,T84.66单链抗体蛋白分子正确表达.T84.66单链抗体可结合KATOⅢ,HGC-27和MKN45,表明这3种细胞表达了特异性肿瘤抗原;单链抗体不能结合SGC7901,GC803,BGC823.42例胃癌组织癌胚抗原阳性率早期和进展期分别为55%(6/11)和61%(19/31),在正常胃黏膜组织标本中无表达,二者之间存在显著性差异(P<0.05).结论:KATOⅢ等胃癌细胞系可表达癌胚抗原.后者在胃癌组织表达水平较高,而正常胃组织不表达.

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