Effect of aberrant disulfide bond formation on protein conformation and molecular property of recombinant therapeutics

Zhang, Lin; Moo-Young, Murray; Chou, C. Perry

doi:10.1351/PAC-CON-09-01-06

Pure Appl. Chem., 2010, Vol. 82, No. 1, pp. 149-159

http://dx.doi.org/10.1351/PAC-CON-09-01-06

Published online 2010-01-03

Effect of aberrant disulfide bond formation on protein conformation and molecular property of recombinant therapeutics

Lin Zhang, Murray Moo-Young and C. Perry Chou*

Department of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada

Abstract: As a comparative study, the extracytoplasmic domain of human CD83 (hCD83ext) was expressed as a glutathione-S-transferase (GST) fusion in two Escherichia coli B strains, i.e., BL21 and Origami B, respectively, with a reductive and oxidative cytoplasm. The final therapeutic products of hCD83ext produced by the two expression hosts exhibited significant differences in protein conformation and molecular property, which presumably resulted from different disulfide patterns. The study highlights the importance of developing proper host/vector systems and biomanufacturing conditions for the production of recombinant therapeutic proteins with a consistent product quality.