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Renaturation, Purification and Characterization of Recombinant Fab-Fragments Produced in Escherichia coli

Bio/Technology volume 9, pages 157–162 (1991)Cite this article

Abstract

Cytoplasmatic expression of murine antibody chains in Escherichia coli results in the formation of insoluble and inactive protein aggregates (inclusion bodies). By systematic variation of the parameters influencing the folding, formation of disul-fide bonds and association of the constituent polypeptide chains, we have designed a renaturation procedure allowing the production of microbially expressed Fab-fragments at yields up to 40 percent of the total amount of recombinant protein. The strategy of optimization is generally applicable for disulnde containing proteins produced as inclusion bodies in bacteria. The purified recombinant antibody fragments obtained are identical with the native murine Fab in all functional and phys-icochemical parameters tested.

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Author notes

  1. Rainer Rudolph: Corresponding author.

Authors and Affiliations

  1. Universität Regensburg, Institut für Biophysik and Physikalische Biochemie, Postfach, D-8400, Regensburg, FRG

    Johannes Buchner

  2. Boehringer Mannheim GmbH, Biochemical Research Center, Nonnenwald 2, D-8122, Penzberg, FRG

    Rainer Rudolph

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Buchner, J., Rudolph, R. Renaturation, Purification and Characterization of Recombinant Fab-Fragments Produced in Escherichia coli. Nat Biotechnol 9, 157–162 (1991). https://doi.org/10.1038/nbt0291-157

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