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Gewählte Publikation:

Wozniak-Knopp, G; Stadlmann, J; Rüker, F; .
(2012): Stabilisation of the Fc fragment of human IgG1 by engineered intradomain disulfide bonds.
PLoS One. 2012; 7(1):e30083 FullText FullText_BOKU

Abstract:
We report the stabilization of the human IgG1 Fc fragment by engineered intradomain disulfide bonds. One of these bonds, which connects the N-terminus of the CH3 domain with the F-strand, led to an increase of the melting temperature of this domain by 10 degrees C as compared to the CH3 domain in the context of the wild-type Fc region. Another engineered disulfide bond, which connects the BC loop of the CH3 domain with the D-strand, resulted in an increase of T-m of 5 degrees C. Combined in one molecule, both intradomain disulfide bonds led to an increase of the T-m of about 15 degrees C. All of these mutations had no impact on the thermal stability of the CH2 domain. Importantly, the binding of neonatal Fc receptor was also not influenced by the mutations. Overall, the stabilized CH3 domains described in this report provide an excellent basic scaffold for the engineering of Fc fragments for antigen-binding or other desired additional or improved properties. Additionally, we have introduced the intradomain disulfide bonds into an IgG Fc fragment engineered in C-terminal loops of the CH3 domain for binding to Her2/neu, and observed an increase of the T-m of the CH3 domain for 7.5 degrees C for CysP4, 15.5 degrees C for CysP2 and 19 degrees C for the CysP2 and CysP4 disulfide bonds combined in one molecule.
Autor*innen der BOKU Wien:
Rüker Florian
Stadlmann Johannes
Wozniak-Knopp Gordana
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Amino Acid Sequence
Binding Sites/genetics
Calorimetry, Differential Scanning
Circular Dichroism
Disulfides/chemistry*
Humans
Immunoglobulin Fc Fragments/chemistry*
Immunoglobulin Fc Fragments/genetics
Immunoglobulin Fc Fragments/metabolism
Immunoglobulin G/chemistry*
Immunoglobulin G/genetics
Immunoglobulin G/metabolism
Mass Spectrometry
Models, Molecular
Molecular Sequence Data
Mutation
Protein Binding
Protein Engineering/methods*
Protein Stability
Protein Structure, Tertiary
Protein Unfolding
Receptor, ErbB-2/chemistry
Receptor, ErbB-2/metabolism
Temperature
Transition Temperature



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