Title | Disulfide bond introduction for general stabilization of immunoglobulin heavy-chain variable domains. |
Publication Type | Journal Article |
Year of Publication | 2008 |
Authors | Saerens, D., K. Conrath, J. Govaert, and S. Muyldermans |
Journal | J Mol Biol |
Volume | 377 |
Issue | 2 |
Pagination | 478-88 |
Date Published | 2008 Mar 21 |
ISSN | 1089-8638 |
Keywords | Amino 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. |
DOI | 10.1016/j.jmb.2008.01.022 |
Alternate Journal | J. Mol. Biol. |
PubMed ID | 18262543 |
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