Efficient capture of monoclonal antibody from cell culture supernatant using protein A media contained in a cuboid packed-bed device

被引:4
作者
Chen, Guoqiang [1 ]
Gerrior, Alisha [1 ]
Durocher, Yves [2 ,3 ]
Ghosh, Raja [1 ]
机构
[1] McMaster Univ, Dept Chem Engn, 1280 Main St West, Hamilton, ON L8S 4L7, Canada
[2] Natl Res Council Canada, Montreal, PQ H4P 2R2, Canada
[3] Univ Montreal, Fac Med, Dept Biochim & Med Mol, Montreal, PQ H3C 3J7, Canada
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2019年 / 1134卷
基金
加拿大自然科学与工程研究理事会;
关键词
Column; Cuboid packed-bed; Chromatography; Capture; Monoclonal antibody; Protein A; Affinity chromatography; CHROMATOGRAPHY; COLUMN; PERFORMANCE; TECHNOLOGY; SEPARATION; RECOVERY; AFFINITY; TRENDS;
D O I
10.1016/j.jchromb.2019.121853
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A box-shaped or cuboid packed-bed device was used for monoclonal antibody (mAb) separation using protein A affinity chromatography. The separation efficiency of the device was compared with an equivalent column i.e. packed with same resin, and having identical bed height and bed volume. The protein A media packed cuboid device had a larger number of theoretical plates than its equivalent column, e.g. 8750/m as opposed to about 4700/m at a flow rate of 0.5 mL/min. In rnAb purification experiments, the impurity flow-through and eluted mAb peaks were shaper with the cuboid device. This implied that the effective separation time and buffer consumption with this device was lower, the purified mAb pooled volume was smaller, and the mAb concentration in the pooled volume was greater. Equivalent separation efficiency could be obtained with the cuboid device using higher flow rates than that used with the column. For instance, elution peaks equivalent to those obtainable by the column could be obtained at a 5 times greater flow rate using the cuboid device. The results discussed in this paper clearly demonstrate the potential for improving the efficiency of protein A affinity chromatography based mAb purification by using a cuboid packed-bed device.
引用
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页数:7
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