Current Proteomics

Bernd Rehm  
Institute of Molecular BioSciences Massey University
Private Bag 11222
Palmerston North
New Zealand

Back

Improved Expression of Single-Chain Fragment Variable Antibodies Devoid of Leader Peptides in the Cytoplasm

Author(s): Nur Faezee Ismail, Soo Khim Chan, Gee Jun Tye, Yee Siew Choong, Theam Soon Lim.

Graphical Abstract:


Abstract:

Single-chain variable fragment (scFv) are commonly expressed and translocated into the periplasmic layer with the aid of a leader peptide. This process allows for proper protein folding with the formation of disulfide bonds. The natural oxidative environment of the periplasm promotes disulfide bond formation compared to the highly reductive environment of the cytoplasm. Thus, a leader peptide is vital for the translocation process with different leader peptides being reported. However this method usually results in lower yields and often incomplete translocation of scFv. In this study, we compared the expression of two scFvs using different cell types without a leader peptide based on yield and functionality. We found that scFv expression without leader peptides in the cytoplasm using SHuffle® T7 Escherichia coli K-12 strain resulted in highest expression yields with preserved functionality. The SHuffle® T7 host is a suitable alternative for high-yield production of functional scFv devoid of leader peptides.

Keywords: Disulfide bond, Peptide leader, scFv, SHuffle® T7, Periplasmic protein translocation, Cytoplasmic protein folding, Protein expression.

Order Reprints Order Eprints Rights & PermissionsPrintExport

Article Details

VOLUME: 12
ISSUE: 2
Year: 2015
Page: [117 - 123]
Pages: 7
DOI: 10.2174/157016461202150903114319
Price: $58