论文部分内容阅读
从抗ＨＩＶ１ 整合酶（ｉｎｔｅｇｒａｓｅ ，ＩＮ） 单链抗体（ｓｉｎｇｌｅ ｃｈａｉｎ ｆｒａｇ ｍｅｎｔ ｖａｒｉａｂｌｅ ，ＳｃＦｖ） 的ＨＢ２１５１ 表达菌中选取３ 个克隆，进行ＳｃＦｖ 的可溶性表达与纯化，并观察ＳｃＦｖ 对ＩＮ 体外生物学活性的影响。结果显示，在ＳｃＦｖ ：ＩＮ摩尔浓度为８ ∶１ 时，３ 个克隆的ＳｃＦｖ 可明显抑制ＩＮ 的３′端加工、链转移及去整合活性； 而相同浓度的ＢＳＡ 及抗人交联纤维的ＳｃＦｖ 对ＩＮ 活性无明显影响。测序结果表明，此３ 个克隆的ＳｃＦｖ 基因序列完全相同，符合鼠抗体可变区序列。完全抑制３′端加工及去整合反应所需ＳｃＦｖ 的浓度高于链转移反应。研究结果提示，抗ＩＮ 的ＳｃＦｖ 可抑制ＩＮ 的体外生物学活性，此结论为进一步研究此类ＳｃＦｖ 对ＨＩＶ 在细胞内复制的影响及其机制奠定了基础。
Three clones were selected from the HB2151 expressing bacterium of anti-HIV-1 integrase (IN) single chain frag ment variable (ScFv) for the soluble expression and purification of ScFv. The effects of ScFv on in vitro biology Effect of activity. The results showed that ScFv of 3 clones could significantly inhibit 3’-end processing, chain transfer and de-integrative activity of IN when the molar concentration of ScFv: IN was 8: 1. However, ScFv of the same concentration of BSA and anti-human crosslinked fiber No significant effect on IN activity. The sequencing results showed that the sequences of the ScFv genes of the three clones were exactly the same and corresponded to the variable regions of the murine antibody. The concentration of ScFv needed to completely inhibit the 3 ’end processing and de-integration reaction is higher than that of the chain transfer reaction. The results suggest that anti-IN ScFv can inhibit in vitro biological activity of IN, the conclusion for further study of such ScFv on HIV replication in the intracellular and its mechanism laid the foundation.