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. Author manuscript; available in PMC: 2012 Mar 1.

Published in final edited form as: Metab Eng. 2010 Dec 3;13(2):241–251. doi: 10.1016/j.ymben.2010.11.002

(A) Yield of assembled 26.10 IgG in different combinations of strain, heavy chain TIR sequence, and DsbA co-expression determined by Western blotting and ELISA (bottom). WT: E. coli JUDE-1 cells. (B) Production of full-length antibody for different IgG variants expressed either in JUDE-1 cells carrying the original pMAZ360 vector or in TM4 carrying the pMAZ360 vector containing the engineered heavy chain TIR sequence. Experiments were carried out in triplicate and the error bars correspond to one standard deviation from the mean values. (C) Western blot analysis of the production of full-length antibody for different IgG variants expressed either in JUDE-1 or TM4 cells carrying pMAZ360 vector with the original heavy chain TIR sequence. All lanes were loaded with total protein corresponding to the same number of cells as judged by OD₆₀₀. (D) SDS-PAGE analysis of 26.10 IgG following protein A affinity chromatography of cell lysates from either JUDE-1 cells carrying the pMAZ360-26.10-IgG vector with the original heavy chain TIR sequence or TM4 cells carrying the pMAZ360-26.10-IgG vector with the engineered heavy chain TIR sequence. MW: molecular mass in kDa.

(A) Yield of assembled 26.10 IgG in different combinations of strain, heavy chain TIR sequence, and DsbA co-expression determined by Western blotting and ELISA (bottom). WT: E. coli JUDE-1 cells. (B) Production of full-length antibody for different IgG variants expressed either in JUDE-1 cells carrying the original pMAZ360 vector or in TM4 carrying the pMAZ360 vector containing the engineered heavy chain TIR sequence. Experiments were carried out in triplicate and the error bars correspond to one standard deviation from the mean values. (C) Western blot analysis of the production of full-length antibody for different IgG variants expressed either in JUDE-1 or TM4 cells carrying pMAZ360 vector with the original heavy chain TIR sequence. All lanes were loaded with total protein corresponding to the same number of cells as judged by OD₆₀₀. (D) SDS-PAGE analysis of 26.10 IgG following protein A affinity chromatography of cell lysates from either JUDE-1 cells carrying the pMAZ360-26.10-IgG vector with the original heavy chain TIR sequence or TM4 cells carrying the pMAZ360-26.10-IgG vector with the engineered heavy chain TIR sequence. MW: molecular mass in kDa.