| 梭曼单链抗体的基因克隆和表达 |
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| 引用本文： | 周永新 荣康泰. 梭曼单链抗体的基因克隆和表达[J]. 中国药理学与毒理学杂志, 1996, 10(4): 272-276 |
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| 作者姓名： | 周永新 荣康泰 |
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| 作者单位： | 毒物药物研究所 |
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| 摘 要： | 成功地扩增了有机磷毒剂梭曼类似物１－羟基－１－对胺基苯基甲膦酸单频哪基醇酯与血蓝蛋白结合物免疫小鼠脾细胞中抗体重链和轻链基因片段，构建了单链抗体可变区基因（ＶＨ－ｌｉｎｋｅｒ－ＶＬＤＮＡ），并将该基因克隆到噬菌体表达载体ｐＣＡＮＴＡＢ５Ｅ的外壳蛋白ｇ３ｐ基因中，使单链抗体以融合蛋白的形式在Ｅ．ｃｏｌｉＴＧ１中表达在噬菌体表面，构成噬菌体抗体库．通过免疫亲和淘洗和竞争抑制酶联免疫吸附测定法筛选，得到１２株克隆，其分泌的单链抗体与梭曼有结合活性．
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| 关 键 词： | 梭曼；抗体.单链；噬菌体；抗体库；克隆.分子；基因表达 |
Molecularcloningandexpresionofanti-somansinglechainFvantibodies |
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| ZHOUYong-Xin,RONGKang-Tai. Molecularcloningandexpresionofanti-somansinglechainFvantibodies[J]. Chinese Journal of Pharmacology and Toxicology, 1996, 10(4): 272-276 |
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| Authors: | ZHOUYong-Xin RONGKang-Tai |
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| Abstract: | RecombinantDNAandgeneamplifica-tiontechnologyhasmadeitpossibletocloneantibodygenesinbacteriaandtoproduceantibodiesquicklyinbacterialcultures.TheafinityandstabilityofsinglechainFvantibodies(ScFv)aregeneralycomparabletothoseofthenativeantibodies.Furthermore,ScFvareespecialyfavorabletotheimmuno-therapeutictreat-mentforthechemicalwarfareagentspoisoningastheyareonlyonesixthofintactantibodyinmolecularweight.Thispaperfirstdescribedthemolecularcloningandexpressionoftheanti-somanScFv.Usingpoly-merasechainreaction,immuno-globulinvariableheavy(VH)andlight(VL)regiongeneswereamplifiedfromthespleencelsofaBALB/cmouseimmunizedwiththeconjugateofpinacolyl1-p-aminophenyl-1-hydroxy-methylpho-sphonateandLimuluspolyphemushemocyanin(P4-LPH).TheVH-linker-VLgeneswereconstructed,thenclonedintoaphagemidvectorpCANTAB5Eandexpressedasafusionwithaphageprotein.Thephage-displayedantibodieswhichwerecapableofbindingsomanwereselectedbypanningagainsttheP4-BSAconjugateanddetectedbyacom-petitiveinhibitionenzymeimmunoassay.TwelvecolonieswereobtainedwhichproducedsolubleScFvan-tibodiescapableofbindingsoman. |
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| Keywords: | soman antibody singlechainFv phagedisplay antibodylibrary cloning molecular geneexpression |
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