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High-level production of a single chain antibody against anthrax toxin in Escherichia coli by high cell density cultivation

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Abstract

Previously, we isolated the M18 scFv, which is an affinity matured antibody against the anthrax toxin PA, and observed that its single chain antibody (scAb) form (M18 scAb) exhibited superior stability compared to the scFv. Here, we report high cell density cultivations for preparative scale production of M18 scAb in a 3.5 L fermenter. Briefly, a pH–stat feeding strategy was employed in fed-batch cultivation, and four different cell densities (OD600 of 40, 80, 120, and 150) were examined for the induction of scAb gene expression. Among the four cell densities investigated, lower cell densities (OD600 of 40) showed higher post-induction cell growth and production yields (665 mg/L of scAb). Even though lower solubility (51%) of scAb was achieved at lower cell density (OD600 of 40), monomeric scAb could be purified with high purity (>95%) using simple purification procedures. The purified scAb from high cell density cultures showed biological activity equivalent to that of scAb purified from shake flask cultivation.

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Acknowledgments

We would like to thank Dr. George Georgiou and Dr. Brent L. Iverson (University of Texas at Austin) for very helpful comments on research and manuscript preparation. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (Grant No. 2010-0011216).

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, KAIST, Daejeon, 305-701, Republic of Korea

    Ki Jun Jeong

  2. Institute for Cellular and Molecular Biology, University of Texas, Austin, TX, 78712, USA

    Mridula Rani

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  1. Ki Jun Jeong
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  2. Mridula Rani
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Correspondence to Ki Jun Jeong.

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Jeong, K.J., Rani, M. High-level production of a single chain antibody against anthrax toxin in Escherichia coli by high cell density cultivation. Bioprocess Biosyst Eng 34, 811–817 (2011). https://doi.org/10.1007/s00449-011-0531-1

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  • DOI: https://doi.org/10.1007/s00449-011-0531-1

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