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Platform development for expression and purification of stable isotope labeled monoclonal antibodies in Escherichia coli

Published

Author(s)

Prasad T. Reddy, Robert G. Brinson, James T. Hoopes, Colleen McClung, Na Ke, Lila Kashi, Mehmet Berkmen

Abstract

Proteins labeled with stable isotopes play an important role in structural and biophysical studies including nuclear magnetic resonance, small angle neutron scattering, neutron reflectometry, quantitative mass spec and more. The most common and cost-efficient methods to labeled proteins is an expression and purification in Escherichia coli growing in minimal medium. Monoclonal antibodies (mAbs) are fast growing segment of the pharmaceutical industries and thus there is a growing interest to study the structure, stability and dynamics of mAbs. However, to date, no study on the expression and purification of mAbs from E. coli growing in minimal medium was reported. Here we report, for the first time, the expression and purification of a labeled mAb expressed and purified from E. coli. It is shown that the unlabeled mAb and the mAb labeled with 13C, 15N, 2H or the triple labeled with 13C, 15N and 2H are well folded and are similar to the same mAb expressed and purified in eukaryotic cells. The implication of these observation to the pharmaceutical industry and the future studies on the structure, biophysical, and biochemical properties of mAb is discussed.
Citation
mAbs
Volume
10
Issue
7

Keywords

protein production, E. coli, isotope labeling, mAb

Citation

Reddy, P. , Brinson, R. , , J. , McClung, C. , Ke, N. , Kashi, L. and Berkmen, M. (2018), Platform development for expression and purification of stable isotope labeled monoclonal antibodies in Escherichia coli, mAbs, [online], https://doi.org/10.1080/19420862.2018.1496879 (Accessed June 22, 2021)
Created October 1, 2018, Updated February 13, 2020