超滤分离蛋白质过程快速优化新技术及其应用

A novel methodology for the rapid process optimization of protein ultrafiltration and its application

介绍了蛋白质超滤分离快速优化新技术、新方法,包括脉冲进样技术,载体相超滤技术、参数连续变化超滤技术以及在这些技术基础上建立的超滤分离快速优化新方法,克服了常规蛋白质超滤分离过程优化中存在的实验工作量大、蛋白质消耗多、费时以及费用高等缺点。利用该方法对以溶菌酶和卵清蛋白为模型蛋白质的混合体系进行了研究,筛选出合适的超滤膜,优化了分离条件.结果表明,在本研究实验条件范围内,30 kDa Biomax膜要优于30 kDaYM膜.利用30 kDa Biomax膜对溶菌酶和卵清蛋白的超滤分离可取得较好的效果,采用载体相超滤技术经240 min后透过液中溶菌酶的纯度>90%,回收率>92%.

A novel methodology, which involves the application and combination of several newly developed techniques such as pulsed sample injection technique, carrier phase ultrafiltration and parameter scanning ultrafiltration, was introduced in this paper. With this methodology, the fractionation of the binary mixture of lysozyme and ovalbumin was performed with 30 kDa Biomax and YM membranes, respectively, and rapid process optimization realized. The experimental results showed that under the operating conditions examined, the 30 kDa Biomax membrane was superior to the 30 kDa YM membrane in the ffactionation of the binary protein mixture. With the 30 kDa Biomax membrane, lysozyme with the purity of more than 90% could be obtained in the permeate, and its recovery was more than 92% after 240 rain operation using carrier phase ultrafiltration.