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Antibody Engineering pp 181–189Cite as

Disulfide-Stabilized Fv Fragments

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Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

The design and generation of disulfide-stabilized Fv fragments (dsFv’s) addresses stability and aggregation problems that are frequently associated with single-chain Fvs. VH and VL of dsFv’s are connected by an interdomain disulfide bond. To generate such molecules, one amino acid each in the framework region of in VH (at position 44) and VL (at position 100) are mutated to a cysteine, which in turn form a stable interchain disulfide bond. The resulting dsFv’s (no linker peptide) or scdsFv (linker as well as interchain disulfide bond) can be easily produced in various expression systems. Disulfide-stabilized Fv’s solve most problems that are frequently associated with Fvs or scFvs; they are very stable and in most instances show full antigen binding activity.

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Abbreviations

IPTG:

Isopropyl-β-d-thiogalactopyranoside

scFv:

Single-chain Fv

dsFv:

Disulfide-stabilized Fv

VH and VL :

Variable region of heavy or light-chain

GuCl:

Guanidine chloride

DTE:

Dithioerythritol

IB:

Inclusion body

IG:

Immunoglobulin

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Author information

Authors and Affiliations

  1. Roche Pharma Research, Biologics R&D, Nonnenwald 2, D-82377, Penzberg, Germany

    Ulrich Brinkmann

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  1. Ulrich Brinkmann
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Correspondence to Ulrich Brinkmann .

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Editors and Affiliations

  1. (Biomedical Engineering) Institut für Zellbiologie und Immunologie, Universität Stuttgart, Allmandring 31, 70569, Stuttgart, Germany

    Roland Kontermann

  2. Technische Universität Braunschweig, Institut für Biochemie und Biotechnologie, Spielmannstraße 7, 38106, Braunschweig, Germany

    Stefan Dübel

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Brinkmann, U. (2010). Disulfide-Stabilized Fv Fragments. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-01147-4_14

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  • DOI: https://doi.org/10.1007/978-3-642-01147-4_14

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-01146-7

  • Online ISBN: 978-3-642-01147-4

  • eBook Packages: Springer Protocols

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