Abstract
The crystal structures of the murine BR96 Fab and its human chimera have been determined in complex with the nonoate methyl ester derivative of Lewis Y (nLey) at 2.8 A and 2.5 A resolution, respectively. BR96 binds the carbohydrate in a large pocket which is formed by residues of all CDR loops except L2. The binding of the carbohydrate is mediated predominantly by aromatic residues in BR96. Analysis of the structure suggests that BR96 is capable of recognizing a structure larger than the Le(y) tetrasaccharide, providing a possible explanation for its high tumour selectivity. The structure provides a rationale for mutagenesis experiments that have resulted in BR96 CDR loop mutants with increased affinity for nLey and/or tumour cells.
MeSH terms
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Animals
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Antibodies, Monoclonal / chemistry*
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Antibodies, Monoclonal / genetics
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Antibodies, Monoclonal / immunology
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Antibodies, Neoplasm / chemistry*
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Antibodies, Neoplasm / immunology
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Antibodies, Neoplasm / therapeutic use
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Binding Sites, Antibody / physiology
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Carbohydrate Sequence
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Crystallography, X-Ray
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Humans
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Immunoglobulin Fab Fragments / chemistry
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Immunoglobulin Fab Fragments / immunology
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Lewis Blood Group Antigens / chemistry*
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Lewis Blood Group Antigens / immunology
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Mice
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Models, Molecular
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Molecular Sequence Data
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Molecular Structure
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Mutation
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Neoplasms / immunology
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Neoplasms / therapy
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Protein Conformation
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Recombinant Fusion Proteins / chemistry*
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Recombinant Fusion Proteins / immunology
Substances
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Antibodies, Monoclonal
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Antibodies, Neoplasm
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Immunoglobulin Fab Fragments
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Lewis Blood Group Antigens
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Lewis Y antigen
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Recombinant Fusion Proteins