4S1D

Structure of IgG1 Fab fragment in complex with Biotincytidinamide

4S1D の概要

• エントリーDOI: 10.2210/pdb4s1d/pdb
• 分子名称: MAB M33 FAB FRAGMENT, heavy chain, MAB M33 FAB FRAGMENT, light chain, N~6~-{5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanoyl}-L-lysinamide, ... (5 entities in total)
• 機能のキーワード: antibody, fab fragment, hapten, mab_m33_fab_fragment, immune system
• 由来する生物種: Mus musculus (mouse); 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 191160.23

構造登録者

Dengl, S.,Hoffmann, E.,Grote, M.,Wagner, C.,Mundigl, O.,Georges, G.,Theorey, I.,Stubenrauch, K.-G.,Bujotzek, A.,Josel, H.-P.,Dziadek, S.,Benz, J.,Brinkmann, U. (登録日: 2015-01-13, 公開日: 2015-03-04, 最終更新日: 2024-10-09)

主引用文献

Dengl, S.,Hoffmann, E.,Grote, M.,Wagner, C.,Mundigl, O.,Georges, G.,Thorey, I.,Stubenrauch, K.G.,Bujotzek, A.,Josel, H.P.,Dziadek, S.,Benz, J.,Brinkmann, U.
Hapten-directed spontaneous disulfide shuffling: a universal technology for site-directed covalent coupling of payloads to antibodies.
Faseb J., 29:1763-1779, 2015

PubMed Abstract: Humanized hapten-binding IgGs were designed with an accessible cysteine close to their binding pockets, for specific covalent payload attachment. Individual analyses of known structures of digoxigenin (Dig)- and fluorescein (Fluo) binding antibodies and a new structure of a biotin (Biot)-binder, revealed a "universal" coupling position (52(+2)) in proximity to binding pockets but without contributing to hapten interactions. Payloads that carry a free thiol are positioned on the antibody and covalently linked to it via disulfides. Covalent coupling is achieved and driven toward complete (95-100%) payload occupancy by spontaneous redox shuffling between antibody and payload. Attachment at the universal position works with different haptens, antibodies, and payloads. Examples are the haptens Fluo, Dig, and Biot combined with various fluorescent or peptidic payloads. Disulfide-bonded covalent antibody-payload complexes do not dissociate in vitro and in vivo. Coupling requires the designed cysteine and matching payload thiol because payload or antibody without the Cys/thiol are not linked (<5% nonspecific coupling). Hapten-mediated positioning is necessary as hapten-thiol-payload is only coupled to antibodies that bind matching haptens. Covalent complexes are more stable in vivo than noncovalent counterparts because digoxigeninylated or biotinylated fluorescent payloads without disulfide-linkage are cleared more rapidly in mice (approximately 50% reduced 48 hour serum levels) compared with their covalently linked counterparts. The coupling technology is applicable to many haptens and hapten binding antibodies (confirmed by automated analyses of the structures of 140 additional hapten binding antibodies) and can be applied to modulate the pharmacokinetics of small compounds or peptides. It is also suitable to link payloads in a reduction-releasable manner to tumor- or tissue-targeting delivery vehicles.

PubMed: 25670234
DOI: 10.1096/fj.14-263665

実験手法

X-RAY DIFFRACTION (2.5 Å)