Increased production and recovery of secreted foreign proteins from plant cell cultures using an affinity chromatography bioreactor
- نوع فایل : کتاب
- زبان : انگلیسی
- مؤلف : Eddie James, David R. Mills, James M. Lee
- چاپ و سال / کشور: 2002
Description
Although plant cell cultures can be used to produce a variety of proteins, the instability and degradation of secreted proteins hinders product recovery from culture medium. To solve this problem, an affinity chromatography bioreactor (ACBR) scheme was developed for the simultaneous production and purification of foreign proteins from genetically modified plant suspension cultures. Two model products, the heavy chain of a mouse monoclonal antibody (HC MAb) and 6-his tagged human granulocyte macrophage colony-stimulating factor (GM-CSF), were produced in a modified shake flask and separated using protein G and iminodiacetic acid metal affinity resins, respectively. Results showed that the ACBR scheme increased the production of HC MAb by as much as eight-fold and GM-CSF by more than two-fold compared with batch yields. Product recovery was optimized by varying the aeration method, flow rate, pH, and resin type for the experimental system. These results suggest that in situ protein removal from plant cell media using the ACBR strategy can isolate the product from degrading influences and remove the protein from product inhibition pathways. © 2002 Elsevier Science B.V. All rights reserved.
Biochemical Engineering Journal 12 (2002) 205–213 Received 2 July 2001; accepted after revision 16 May 2002
