title: Anti-c-Met antibodies recognising a temperature sensitive epitope, inhibit cell growth.
abstract: c-Met is a tyrosine receptor kinase which is activated by its ligand, the hepatocyte growth factor. Activation of c-Met leads to a wide spectrum of biological activities such as motility, angiogenesis, morphogenesis, cell survival and cell regeneration. c-Met is abnormally activated in many tumour types. Aberrant c-Met activation was found to induce tumour development, tumour cell migration and invasion, and the worst and final step in cancer progression, metastasis. In addition, c-Met activation in cells was also shown to confer resistance to apoptosis induced by UV damage or chemotherapeutic drugs. This study describes the development of monoclonal antibodies against c-Met as therapeutic molecules in cancer treatment/diagnostics. A panel of c-Met monoclonal antibodies was developed and characterised by epitope mapping, Western blotting, immunoprecipitation, agonist/antagonist effect in cell scatter assays and for their ability to recognise native c-Met by flow cytometry. We refer to these antibodies as Specifically Engaging Extracellular c-Met (seeMet). seeMet 2 and 13 bound strongly to native c-Met in flow cytometry and reduced SNU-5 cell growth. Interestingly, seeMet 2 binding was strongly reduced at 4oC when compared to 37oC. Detail mapping of the seeMet 2 epitope indicated a cryptic binding site hidden within the c-Met α-chain.
authors.affiliation: p53 Laboratory, 8A Biomedical Grove, Immunos #06-06, Singapore, Singapore.
authors.forename: Julin S
authors.initials: JS
authors.lastname: Wong
authors.forename: Emma
authors.initials: E
authors.lastname: Warbrick
authors.forename: Borek
authors.initials: B
authors.lastname: Vojtesk
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Hill
authors.forename: David P
authors.initials: DP
authors.lastname: Lane
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D048708:Cell Growth Processes / Q000187:drug effects; D045744:Cell Line, Tumor; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D019859:Proto-Oncogene Proteins c-met / Q000037:antagonists & inhibitors* / Q000276:immunology*; D015398:Signal Transduction; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D019859:Proto-Oncogene Proteins c-met