Disulfide bond introduction for general stabilization of immunoglobulin heavy-chain variable domains.

TitleDisulfide bond introduction for general stabilization of immunoglobulin heavy-chain variable domains.
Publication TypeJournal Article
Year of Publication2008
AuthorsSaerens, D., K. Conrath, J. Govaert, and S. Muyldermans
JournalJ Mol Biol
Volume377
Issue2
Pagination478-88
Date Published2008 Mar 21
ISSN1089-8638
KeywordsAmino Acid Sequence, Cysteine, Disulfides, Immunoglobulin Heavy Chains, Immunoglobulin Variable Region, Models, Molecular, Molecular Sequence Data, Mutation, Nanostructures, Protein Folding, Protein Structure, Tertiary, Sequence Alignment
Abstract

Several antibody fragment engineering techniques aim at intrinsic stability enhancement, but are not applied in a truly generic way. Here, a strategy is proposed whereby consistent gain in stability is accomplished by introducing a specific disulfide bond between two opposite beta-strands in the hydrophobic core of the immunoglobulin heavy-chain variable domain of heavy-chain antibodies (Nanobody). Besides the rational design of a disulfide bond between residues 39 and 87, a Nanobody harboring an extra naturally occurring cystine between residues 54 and 78 was compared to an equivalent Nanobody without that cystine. Both novel disulfide cross-links were introduced in several Nanobodies in various combinations. Interestingly, only the extra naturally occurring cystine consistently increased the conformational and thermal stabilities of wild-type Nanobodies without affecting antigen binding.

DOI10.1016/j.jmb.2008.01.022
Alternate JournalJ. Mol. Biol.
PubMed ID18262543