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An intrinsically stable antibody scFv fragment can tolerate the loss of both disulfide bonds and fold correctly.
Journal article

An intrinsically stable antibody scFv fragment can tolerate the loss of both disulfide bonds and fold correctly.

  • 1998-06-24
Published in:
  • FEBS letters. - 1998
Amino Acid Substitution
Biosensing Techniques
Chromatography, Gel
Cysteine
Disulfides
Enzyme-Linked Immunosorbent Assay
Immunoglobulin Fragments
Immunoglobulin Heavy Chains
Immunoglobulin Light Chains
Immunoglobulin Variable Region
Point Mutation
Protein Denaturation
Protein Folding
Recombinant Proteins
Thermodynamics
Urea
English A fully functional cysteine-free derivative of the intrinsically stable anti-HER2 scFv fragment hu4D5-8 was generated by replacing the disulfide forming cysteine residues in VH and VL with the amino acid combination valine-alanine in both domains. The antigen binding properties, determined by ELISA and BIAcore measurements, were not affected by removal of the disulfide bonds. The thermodynamic stability of the disulfide-containing scFv of 8.1 kcal/mol is decreased upon complete reduction of both disulfides to 2.7 kcal/mol, while that of the valine-alanine variant is somewhat higher (about 3.8 kcal/ mol). Our results suggest that, in principle, a disulfide-free fully functional derivative of any scFv can be obtained, as long as the corresponding disulfide-containing scFv has a high enough thermodynamic stability.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/55197
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