本文選題：親和微濾 + 親和膜��； 參考：《浙江工業(yè)大學(xué)》2012年碩士論文

【摘要】：目前，抗體已被廣泛應(yīng)用于疾病的診斷、治療以及靶向載體等方面，但是其價(jià)格昂貴，造成其昂貴的主要原因之一在于其純化成本高，雖然目前有許多方法用于純化抗體，但都存在著諸如分辨率不高、處理量不大或分離介質(zhì)昂貴等缺陷，因此開發(fā)更廉價(jià)、更簡便的純化方法十分必要。本課題以抗HCG抗體為模型蛋白，選用廉價(jià)的小分子親和配基（磺胺二甲基嘧啶），并結(jié)合膜分離處理量大的優(yōu)點(diǎn)，進(jìn)行抗體純化新方法的研究，研究取得的結(jié)果如下： 首先，以葡聚糖凝膠微球?yàn)檩d體，磺胺二甲基嘧啶為親和配基，通過環(huán)氧接枝，制備了親和微濾載體，并對制備條件進(jìn)行了優(yōu)化，所制得的載體配基密度為85.5μmol/g（濕），而且重復(fù)利用率高，耐酸堿性及機(jī)械強(qiáng)度良好；將其應(yīng)用于親和微濾分離抗體，最佳吸附pH值為6.0，吸附容量為8.1mg/g（濕），一步分離抗體純度大于95%，回收率69.8%；將其用于柱層析，雖然在酸性條件下洗脫不如蛋白A柱，但在高鹽條件下洗脫效果與蛋白A柱相當(dāng)，活性保持良好。 選用不同的膜基質(zhì)，通過環(huán)氧接枝，制備了磺胺二甲基嘧啶親和膜，并對制備條件進(jìn)行了優(yōu)化，結(jié)果表明，醋酸纖維素膜為膜基質(zhì)的效果較好，最終配基密度為114.6μmol/g，將其應(yīng)用于抗體分離，最佳吸附為pH6.0，，吸附容量為5.7mg/g，一步純化抗體的純度大于95%，回收率37.6%。
[Abstract]:At present, antibody has been widely used in the diagnosis, treatment and targeting vector of disease, but it is expensive, one of the main reasons for its expensive is the high cost of purification, although there are many methods to purify antibody, However, there are some disadvantages such as low resolution, low processing capacity or expensive separation medium. Therefore, it is necessary to develop cheaper and simpler purification methods. In this paper, we use anti-HCG antibody as model protein, select cheap small molecular affinity ligand (sulfadimethylpyrimidine), and combine the advantages of large membrane separation and treatment capacity to study the new method of antibody purification. The results are as follows: Firstly, using dextran gel microspheres as carrier and sulfadimethylpyrimidine as affinity ligand, the affinity microfiltration carrier was prepared by epoxy grafting, and the preparation conditions were optimized. The ligand density of the carrier was 85.5 渭 mol / g (wet), and its reutilization rate was high, acid and alkali resistance and mechanical strength were good, and it was used to isolate antibody by affinity microfiltration, and the ligand density of the carrier was 85.5 渭 mol / g. The optimum adsorption pH value was 6.0, the adsorption capacity was 8.1 mg / g (wet), the purity of one-step separation antibody was greater than 95, and the recovery was 69.8%. It was used in column chromatography, although it was better than protein A column in acid condition, but the elution effect was equivalent to that of protein A column under high salt condition. The activity remains good. The sulfadimethylpyrimidine affinity membrane was prepared by epoxy grafting with different membrane substrates. The preparation conditions were optimized. The results showed that cellulose acetate membrane was a good membrane matrix. The final ligand density was 114.6 渭 mol / g. The best adsorption was pH6.0, the adsorption capacity was 5.7 mg / g, the purity of one-step purified antibody was greater than 95%, and the recovery was 37.6%.
【學(xué)位授予單位】：浙江工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R392

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