title: Engineering and characterisation of chimeric monoclonal antibody 806 (ch806) for targeted immunotherapy of tumours expressing de2-7 EGFR or amplified EGFR.
abstract: We report the generation of a chimeric monoclonal antibody (ch806) with specificity for an epitope on the epidermal growth factor receptor (EGFR) that is different from that targeted by all other anti-EGFR therapies. Ch806 antibody is reactive to both de2-7 and overexpressed wild-type (wt) EGFR but not native EGFR expressed in normal tissues at physiological levels. Ch806 was stably expressed in CHO (DHFR -/-) cells and purified for subsequent characterisation and validated for use in preliminary immunotherapy investigations. Ch806 retained the antigen binding specificity and affinity of the murine parental antibody. Furthermore, ch806 displayed enhanced antibody-dependent cellular cytotoxicity against target cells expressing the 806 antigen in the presence of human effector cells. Ch806 was successfully radiolabelled with both iodine-125 and indium-111 without loss of antigen binding affinity or specificity. The radioimmunoconjugates were stable in the presence of human serum at 37 degrees C for up to 9 days and displayed a terminal half-life (T(1/2beta)) of approximately 78 h in nude mice. Biodistribution studies undertaken in BALB/c nude mice bearing de2-7 EGFR-expressing or amplified EGFR-expressing xenografts revealed that (125)I-labelled ch806 failed to display any significant tumour retention. However, specific and prolonged tumour localisation of (111)In-labelled ch806 was demonstrated with uptake of 31%ID g(-1) and a tumour to blood ratio of 5 : 1 observed at 7 days postinjection. In vivo therapy studies with ch806 demonstrated significant antitumour effects on established de2-7 EGFR xenografts in BALB/c nude mice compared to control, and both murine 806 and the anti-EGFR 528 antibodies. These results support a potential therapeutic role of ch806 in the treatment of suitable EGFR-expressing tumours, and warrants further investigation of the potential of ch806 as a therapeutic agent.
authors.affiliation: Ludwig Institute for Cancer Research, Melbourne Tumour Biology Branch, Level 1, Harold Stokes Building, Austin Hospital, 145-163 Studley Road, Heidelberg 3084, Victoria, Australia.
authors.forename: C
authors.initials: C
authors.lastname: Panousis
authors.forename: V M
authors.initials: VM
authors.lastname: Rayzman
authors.forename: T G
authors.initials: TG
authors.lastname: Johns
authors.forename: C
authors.initials: C
authors.lastname: Renner
authors.forename: Z
authors.initials: Z
authors.lastname: Liu
authors.forename: G
authors.initials: G
authors.lastname: Cartwright
authors.forename: F-T
authors.initials: FT
authors.lastname: Lee
authors.forename: D
authors.initials: D
authors.lastname: Wang
authors.forename: H
authors.initials: H
authors.lastname: Gan
authors.forename: D
authors.initials: D
authors.lastname: Cao
authors.forename: A
authors.initials: A
authors.lastname: Kypridis
authors.forename: F E
authors.initials: FE
authors.lastname: Smyth
authors.forename: M W
authors.initials: MW
authors.lastname: Brechbiel
authors.forename: A W
authors.initials: AW
authors.lastname: Burgess
authors.forename: L J
authors.initials: LJ
authors.lastname: Old
authors.forename: A M
authors.initials: AM
authors.lastname: Scott
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000493:pharmacokinetics / Q000627:therapeutic use*; D016466:CHO Cells; D006224:Cricetinae; D066246:ErbB Receptors / Q000276:immunology*; D005260:Female; D006801:Humans; D007167:Immunotherapy; D051379:Mice; D008807:Mice, Inbred BALB C; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000628:therapy*; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000493:pharmacokinetics / Q000627:therapeutic use*; D014018:Tissue Distribution; D014183:Transplantation, Heterologous
chemical_list: D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D066246:ErbB Receptors