A cleavable self-aggregating tag scheme for the expression and purification of disulfide bonded proteins and peptides

作者：Lin, Zhanglin^*; Jing, Yanyun; Huang, Yuan; Yang, Sha; Chen, Shuyi; Ou, Yingying; Pistolozzi, Marco; Yang, Xiaofeng^*

来源：Chemical Engineering Sciences, 2022, 262: 118052.

DOI：10.1016/j.ces.2022.118052

• Protein expression and purification
• Disulfide bonded proteins and peptides
• Aggregating tag
• Intein
• Escherichia coli

The production of correctly folded, disulfide bonded proteins in high yields remains a technological chal-lenge, especially for the commonly used expression system Escherichia coli. This work provides a stan-dardizable method based on cleavable self-aggregating tag (cSAT) scheme that enables the facile production of a series of traditionally challenging disulfide bonded proteins and peptides. The yields of the six targets were in the range of 3 to 89 mg/L at the shake flask scale. Further scale up of human growth hormone resulted in a yield of 2.6 g/L in a 30-L fermenter. This cSAT scheme allows the correct folding of proteins and peptides in an aggregated form during expression, and can be combined to a stan-dard two-step column purification to achieve high purity and high activity products. This promises a gen-erally applicable platform for the production and purification of proteins and peptides with markedly reduced development time and cost.

• 单位
  南方医科大学