Purification of recombinant human antithrombin III expressed in a goat mammary bioreactor

Cuijie Wang, Yongdong Huang*, Yingjun Kong, Jian Luo, Guifeng Zhang, Dongxu Zhao, Zhiguo Su, Guanghui Ma

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Antithrombin III (AT III) is the most important anti-clotting substance. Recombinant human antithrombin III (rhAT III) expressed in transgenic goat milk attracts more and more attention. Develop an effective purification route for rhAT III is vital to its industrial production. An efficient purification method was developed for the rapid purification of rhAT III by isoelectric precipitation and heparin affinity chromatography. First, casein was effectively removed by isoelectric precipitation. rhAT III was further purified by heparin affinity chromatography. In the process of heparin affinity chromatography, the effects of pH and temperature on the stability of rhAT III were studied, and the effects of operating conditions, elution gradient, flow rate and sample loaded, on the purification efficiency were also studied. Under the optimized conditions, the protein recovery of rhAT III was about 90% with purity over 99%, while its activity recovery was about 50%. Such a purification process is very simple and effective, and it would provide a valuable reference for the further scaling-up of industrial production.

Original languageEnglish
Pages (from-to)1634-1638
Number of pages5
JournalShengwu Gongcheng Xuebao/Chinese Journal of Biotechnology
Volume30
Issue number10
DOIs
Publication statusPublished - 25 Oct 2014

Keywords

  • Heparin affinity chromatography
  • Mammary gland bioreactor
  • Purification
  • Reconstruction human antithrombin III

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