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Improved Bivalent Miniantibodies, with Identical Avidity as Whole Antibodies, Produced by High Cell Density Fermentation of Escherichia coli

Bio/Technology volume 11, pages 1271–1277 (1993)Cite this article

Abstract

The combination of single-chain Fv-fragments (scFv) with a C-terminal, flexible linking region followed by a designed or natural dimerization domain provides a versatile system for targeted association of functional fragments in the periplasmic space of Escherichia coli. For homodimerization in vivo, two scFv fragments with a C-terminal hinge followed by a helix-turn-helix motif form “miniantibodies” with significantly higher avidity than in the case of leucine zipper containing constructs. The favorable design probably results in an antiparallel four-helix bundle and brings the homodimer to the same avidity as the whole IgA antibody, from which the binding site was taken. The molecular weight of the bivalent miniantibody is almost the same as that of a monovalent Fab fragment. We report here a high-cell density fermentation of E. coli producing these miniantibodies and a work-up procedure suitable for large scale production. Without any need of subsequent chemical coupling in vitro, approximately 200 mg/l of functional dimeric miniantibodies can be directly obtained from the E. coli culture.

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Author information

Author notes

  1. Peter Pack

    Present address: MorphoSys, Frankfurter Ring 193a, D-80807, München, Fed. Rep. Germany

  2. Andreas Plückthun: Corresponding author. Address after October 1993: Biochemisches Institut, Universität Zürich, Winterthurerstr. 190, CH-8057 Zürich, Switzerland.

Authors and Affiliations

  1. Max-Planck-Institut für Biochemie, Protein Engineering Group, Am Klopferspitz, D-82152, Martinsried, Fed. Rep. Germany

    Andreas Plückthun

  2. Hans-Knöll-Institut für Naturstoff-Forschung, Beutenbergstr. 11, D-07745, Jena, Fed. Rep. Germany

    Marian Kujau, Volker Schroeckh, Uwe Knüpfer, Rolf Wenderoth & Dieter Riesenberg

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  1. Peter Pack
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Pack, P., Kujau, M., Schroeckh, V. et al. Improved Bivalent Miniantibodies, with Identical Avidity as Whole Antibodies, Produced by High Cell Density Fermentation of Escherichia coli. Nat Biotechnol 11, 1271–1277 (1993). https://doi.org/10.1038/nbt1193-1271

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