Generation of a humanised single chain Fv (Scfv) derived from the monoclonal Eric-1 recognising the human neural cell adhesion molecule.
BACKGROUND
Murine monoclonal antibody (MoAb) ERIC-1 recognises an epitope on the neural cell adhesion molecule (NCAM) whose expression in paediatric and adult tissues is confined mainly to the brain, peripheral nerve, and adrenal medulla. Anti-NCAM antibodies have been used for the treatment and diagnosis of a number of tumours, including neuroblastoma. However, whole antibody exhibits poor penetration into solid tumour deposits and rapid systemic clearance upon repeated administration due to development of a human antimouse antibody (HAMA) response.


METHODS
To overcome these problems, recombinant DNA techniques have been used to humanise and assemble the ERIC-1 immunoglobulin variable heavy (VH) and light (VL) chains into a single chain Fv (scFv).


RESULTS
Three humanised scFv clones were identified which differ from the predicted humanised sequence by occasional amino acid changes, but these maintain the same specificity as the original ERIC-1 MoAb.


CONCLUSIONS
The humanised scFv may prove to be a useful reagents in the treatment and diagnosis of a variety of neuroectodermal tumours and can clearly form a suitable template for the generation of a fully humanised ERIC-1 MoAb.
Whittington
H A
HA
Division of Oncology, University of Bristol, United Kingdom.
Hancock
J
J
Kemshead
J T
JT
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D001667:Binding, Competitive; D015195:Drug Design; D000939:Epitopes / Q000276:immunology; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D019006:Neural Cell Adhesion Molecules / Q000276:immunology*; D009447:Neuroblastoma / Q000276:immunology* / Q000473:pathology; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D019006:Neural Cell Adhesion Molecules; D011993:Recombinant Fusion Proteins