High-resolution purification of a therapeutic PEGylated protein using a cuboid packed-bed device Journal Articles uri icon

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  • PEGylated proteins which are a class of protein-synthetic polymer conjugates that have shown significant promise in the area of biotherapeutics are difficult to purify. A cuboid packed-bed device was used to purify a mono-PEGylated therapeutic protein from impurities such as high molecular weight (HMW) species (e.g., tri- and/or di-PEGylated forms), and low molecular weight (LMW) species such as unreacted protein and polyethylene glycol (or PEG). The separation efficiency of this device was compared with that of an equivalent cylindrical column. The effects of operating conditions such as flow rate, buffer composition, elution gradient, and column loading were systematically compared. An equivalent column with the same bed volume, same resin and same bed height was served as control. In mono-PEGylated protein purifications experiments, the cuboid packed-bed device exhibited sharper peaks and gave better resolution at all conditions examined in this study. The purity of mono-PEGylated protein in the samples collected from the cuboid packed-bed device and the column were comparable, i.e., 98.1% and 97.9% respectively. The recovery of mono-PEGylated protein in the pooled eluate from the cuboid packed-bed device was 31.7% greater than that recovered in the pooled eluate from the column. Therefore, significantly higher recovery of mono-PEGylated protein was obtained with the cuboid packed-bed device while maintaining the same purity specification as obtained with the column.


  • Chen, Guoqiang
  • Umatheva, Umatheny
  • Pagano, John
  • Yu, Deqiang
  • Ghose, Sanchayita
  • Li, Zhengjian
  • Ghosh, Raja

publication date

  • October 2020