中文摘要：

作为蛋白A色谱的重要替代技术，混合模式色谱（mixed-mode chromatography，MMC）具有稳定、廉价和选择性好等优势。本文考察了以Sepharose Fast Flow为基质、4-（咪唑-1-基）苯胺（AN）为配基的新型色谱介质AN SepFF中配基密度对抗体吸附行为和分离性能的影响。研究结果表明，AN SepFF色谱介质在抗体吸附中具有21．2mmol／L的临界配基密度。在临界配基密度以上，抗体在色谱介质上具有较高的吸附容量和很低的解离常数并展现了相当程度的盐耐受性。动态吸附结果进一步表明，抗体的色谱介质中具有与商品化介质相当的动态吸附容量。抗体纯化的结果也表明，在临界配基浓度以上AN SepFF色谱表现出较好的纯化抗体的能力，其抗体产品纯度高于目前商品化的色谱介质：配基密度为55mmol／L时，AN SepFF色谱纯化抗体的收率达到91．9％。由此，AN SepFF色谱介质展现了巨大的应用前景。

英文摘要：

Mixed mode chromatography （MMC） is considered as an important alternative toprotein A chromatography in the capture of antibody based on advantage of chemical stability,cheap price and high selectivity. In this work, a series of novel MMC beads, AN SepFF, with4-（1H-imidazol-l-yl） aniline （AN） ligand was prepared to evaluate the influence of liganddensity on adsorption and chromatographic performance in the purification of antibody. Theresults indicated that there existed a critical ligand density of 21.2mmol/L in antibody adsorptionon AN SepFF adsorbents. Above the critical ligand density, antibody adsorption not only wasrepresented as much higher adsorption capacity and lower dissociation constant, but alsoexhibited great tolerance to salt concentration. Dynamic binding capacity （DBC） for antibodyfurther showed that antibody had a high DBC value equivalent to commercial adsorbents. In thepurification of antibody from rabbit anti-smallpox plasma, the results showed that AN SepFFchromatography had greater performance in antibody purification than commercial MEPHyperCel or MabSelect SuRe chromatography. The yield of antibody reached to 91.9% in ANSepFF chromatography with ligand density of 55mmol/L. Therefore, AN SepFF adsorbents havea powerful potential in the purification of antibody.