化学
色谱法
洗脱
离子色谱法
离子交换
分析化学(期刊)
梯度洗脱
盐(化学)
柱色谱法
单克隆抗体
亲和层析
亲水作用色谱法
高效液相色谱法
作者
Takashi Ishihara,Shuichi Yamamoto
标识
DOI:10.1016/j.chroma.2004.10.040
摘要
Simple methods for the optimization of ion-exchange chromatography of proteins in our previous papers were applied to cation-exchange chromatography purification of monoclonal antibodies (Mab). We carried out linear gradient elution experiments, and obtained the data for the peak salt concentration and the peak width. From these data, the distribution coefficient as a function of salt concentration, and the height equivalent to a theoretical plate (HETP) as a function of mobile phase velocity were calculated. The optimized linear gradient elution conditions were determined based on the relationship between buffer consumption and separation time. The optimal stepwise elution conditions were determined based on the relationship between the distribution coefficient and the salt concentration.
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