Crystal structure of a diabody, a bivalent antibody fragment.
BACKGROUND
Diabodies are dimeric antibody fragments. In each polypeptide, a heavy-chain variable domain (VH) is linked to a light-chain variable domain (VL) but unlike single-chain Fv fragments, each antigen-binding site is formed by pairing of one VH and one VL domain from the two different polypeptides. Diabodies thus have two antigen-binding sites, and can be bispecific. Direct structural evidence is lacking for the connections and dimeric interactions between the two polypeptides of the diabody.


RESULTS
The 2.6 A resolution structure has been determined for a bivalent diabody with a flexible five-residue polypeptide linker between the (amino-terminal) VH and (carboxy-terminal) VL domains. The asymmetric unit of the crystal consists of four polypeptides comprising two diabodies; for one of these polypeptides the linker can be traced between the VH and VL domains. Within each diabody the two associated VH and VL domains make back-to-back interactions through the VH domains, and there is an extensive VL-VL interface between the two diabodies in the asymmetric unit.


CONCLUSIONS
The structure of the diabody is very similar to that which had been predicted by molecular modelling. Diabodies directed against cell-surface antigens should be capable of bringing together two cells, such as in cell-targeted therapy, because the two antigen-binding sites of the diabody are at opposite ends of the molecule and separated by approximately 65 A.
Perisic
O
O
Centre for Protein Engineering, MRC Centre, Cambridge, UK.
Webb
P A
PA
Holliger
P
P
Winter
G
G
Williams
R L
RL
D000595:Amino Acid Sequence; D000818:Animals; D000937:Antigen-Antibody Reactions; D003432:Cross-Linking Reagents / Q000737:chemistry; D018360:Crystallography, X-Ray; D006825:Hybridomas; D007128:Immunoglobulin Fragments / Q000737:chemistry* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D011487:Protein Conformation
D003432:Cross-Linking Reagents; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D046911:Macromolecular Substances