Bacterial Expression, Purification, and Characterization of Single-Chain Antibodies

  • Sergey M. Kipriyanov
Part of the Springer Protocols Handbooks book series (SPH)


In the past few years, some of the limitations of monoclonal antibodies as therapeutic and diagnostic agents have been addressed by genetic engineering. Such an approach is particularly suitable because of the domain structure of the antibody molecule, where functional domains carrying antigen-binding activities (Fabs or Fvs) or effector functions (Fc) can be exchanged between antibodies Fig. 1 A). Smaller antibody-derived molecules include enzymatically produced 50-kDa Fab fragments and engineered 25-kDa single-chain Fv (scFv) consisting of the heavy and light chain variable regions (VH and VL) connected by a flexible 14-24 amino acid long peptide linker (1,2) Fig. 1 B). Compared to IgG molecules, scFv exhibit significantly improved tumor specificity and intratumoral penetration (3-5). However, the rapid blood clearance and monovalent nature of scFv fragments result in considerably lower quantitative tumor retention of these molecules (3,6).


Amersham Pharmacia Biotech Antibody Fragment Glycine Betaine Immobilize Metal Affinity Chromatography scFv Fragment 
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Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Sergey M. Kipriyanov
    • 1
  1. 1.Affimed Therapeutics AGLadenburgGermany

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