Authors: Yong Jae Lee Ki Jun Jeong
Publish Date: 2013/08/14
Volume: 18, Issue: 4, Pages: 751-758
Abstract
In Escherichi coli Secdependent 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 hostvector system for the secretory production of single chain Fv scFv via signal recognition particle SRPdependent pathway instead of Secdependent pathway Use of DsbA signal peptide for SRPdependent 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 coexpression of YidC could significantly improve the solubility of scFv with high expression level For the largescale production fedbatch cultivations with engineered hostvector 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
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