Expression of full-length immunoglobulins in Escherichia coli: rapid and efficient production of aglycosylated antibodies.
Many research and clinical applications require large quantities of full-length antibodies with long circulating half-lives, and production of these complex multi-subunit proteins has in the past been restricted to eukaryotic hosts. In this report, we demonstrate that efficient secretion of heavy and light chains in a favorable ratio leads to the high-level expression and assembly of full-length IgGs in the Escherichia coli periplasm. The technology described offers a rapid, generally applicable and potentially inexpensive method for the production of full-length therapeutic antibodies, as verified by the expression of several humanized IgGs. One E. coli-derived antibody in particular, anti-tissue factor IgG1, has been thoroughly evaluated and has all of the expected properties of an aglycosylated antibody, including tight binding to antigen and the neonatal receptor. As predicted, the protein lacks binding to C1q and the FcgammaRI receptor, making it an ideal candidate for research purposes and therapeutic indications where effector functions are either not required or are actually detrimental. In addition, a limited chimpanzee study suggests that the E. coli-derived IgG1 retains the long circulating half-life of mammalian cell-derived antibodies.
Simmons
Laura C
LC
Department of Molecular Biology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
Reilly
Dorothea
D
Klimowski
Laura
L
Raju
T Shantha
TS
Meng
Gloria
G
Sims
Paul
P
Hong
Kyu
K
Shields
Robert L
RL
Damico
Lisa A
LA
Rancatore
Patricia
P
Yansura
Daniel G
DG
D000818:Animals; D004926:Escherichia coli; D015870:Gene Expression; D005796:Genes; D005822:Genetic Vectors; D006031:Glycosylation; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D007136:Immunoglobulins / Q000235:genetics*; D002679:Pan troglodytes; D013925:Thromboplastin / Q000276:immunology*; D013997:Time Factors
D007074:Immunoglobulin G; D007136:Immunoglobulins; D013925:Thromboplastin