Crystal structure of a chimeric Fab' fragment of an antibody binding tumour cells.
The crystal structure of a chimeric Fab' fragment of a monoclonal antibody is presented. The Fab' comprises the murine light chain and heavy chain variable domains of the carcinoma-binding antibody B72.3 fused to the constant domain of human kappa, and the first constant domain and hinge domain of human gamma 4, respectively. A model for the Fab' has been determined by molecular replacement and refined to a resolution of 3.1 A with an R-factor of 17.6%. The additional residues that distinguish a Fab' from a Fab fragment are seen to be disordered in the crystals. The H3 hypervariable loop is short and adopts a sharp hairpin turn in a conformation that results from an interaction between the lysine side-chain of H93 and the main-chain carbonyl group of H96. The remaining hypervariable loops display conformations similar to those predicted from the canonical structures approach, although loop H2 is apparently displaced by a salt-bridge formed between H55 Asp and the neighbouring H73 Lys. These and other features of the structure likely to be important in grafting the hypervariable loops to an otherwise human framework are discussed.
Brady
R L
RL
Department of Chemistry, University of York, U.K.
Edwards
D J
DJ
Hubbard
R E
RE
Jiang
J S
JS
Lange
G
G
Roberts
S M
SM
Todd
R J
RJ
Adair
J R
JR
Emtage
J S
JS
King
D J
DJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000648:ultrastructure; D000912:Antibodies, Neoplasm / Q000648:ultrastructure*; D002277:Carcinoma / Q000276:immunology; D003461:Crystallography; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000648:ultrastructure*; D051379:Mice; D008969:Molecular Sequence Data; D010315:Particle Accelerators; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D007140:Immunoglobulin Fab Fragments; D011993:Recombinant Fusion Proteins