Enhanced production of antibody fragment via SRP pathway engineering in Escherichia coli

Cited 9 time in webofscience Cited 0 time in scopus
  • Hit : 47
  • Download : 0
In Escherichi coli, Sec-dependent pathway is the major pathway for protein secretion into periplasm, and it has been widely used for the production of antibody fragment. However, in many cases, the production yields of antibody fragments were not satisfactory due to inefficient secretion and low solubility. Here, we have developed the host-vector system for the secretory production of single chain Fv (scFv) via signal recognition particle (SRP)-dependent pathway instead of Sec-dependent pathway. Use of DsbA signal peptide for SRP-dependent pathway allowed more efficient production of scFv compared with Secdependent pathway. To further improve the production yield and solubility of scFv via SRP pathway, the effect of several factors which are closely related to SRP pathway were examined. Among those factors, the co-expression of YidC could significantly improve the solubility of scFv with high expression level. For the large-scale production, fed-batch cultivations with engineered host-vector system were performed and, two different nutrient feeding solutions (complex vs. defined) were examined. When defined feeding solution was supplied, higher production yield (90 mg/L of scFv) could be obtained than complex feeding solution.
Publisher
KOREAN SOC BIOTECHNOLOGY & BIOENGINEERING
Issue Date
2013-07
Language
English
Article Type
Article
Keywords

CELL-DENSITY CULTURE; HUMAN LEPTIN; PROTEIN; EXPRESSION; MEMBRANE; TRANSLOCATION; CULTIVATION; STRAIN

Citation

BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, v.18, no.4, pp.751 - 758

ISSN
1226-8372
DOI
10.1007/s12257-013-0111-0
URI
http://hdl.handle.net/10203/254769
Appears in Collection
CBE-Journal Papers(저널논문)
Files in This Item
There are no files associated with this item.
This item is cited by other documents in WoS
⊙ Detail Information in WoSⓡ Click to see webofscience_button
⊙ Cited 9 items in WoS Click to see citing articles in records_button

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0