亲和层析法纯化小鼠腹水来源的单克隆抗体的实验研究

Studies on Purification Monoclonal Antibody from Mouse Ascites by using Affinity Chromatography

目的建立从小鼠腹水中纯化抗CD28单克隆抗体(mAb)2F5的一步亲和层析方法。方法小鼠mAb腹水样品经离心过滤后,进行亲和层析柱纯化。分别对样品稀释倍数、上样流速、梯度洗脱体积对m A b纯度的影响进行检测。结果样品在稀释4倍以上、上样流速为1.0mL/min以及Gly-HCl(pH 2.7)一步洗脱条件下,可获得纯度大于90%的mAb 2F5,回收率为80%。结论该研究建立的一步亲和层析纯化方法操作简便,所得mAb的纯度和回收率较高。

Objective To develop a one-step affinity chromatography（AC） purification method of anti-CD28 monoclonal antibody（mAb） 2F5 from mouse ascites.Methods After filtrated by centrifugation,the ascites sample was loaded on a Protein G affinity chromatography column and purified by using GIy-HCl（pH 2.7） elution buffer.The effects of sample dilution ratio,flow rate and elution column volume on the purity of antibody obtained were investigated.Results It was shown that mAb 2F5 with a purity of over 90% could be achieved by using sample dilution above 4 folds,flow rate at 1.0mL/min and one step elution.The total recovery rate of the mAb was 80%.Conclusion The established one-step affinity chromatography purification method was simple and suitable for purification of mAb 2F5.