title: In vivo tumor delivery of a recombinant single chain Fv::tumor necrosis factor-alpha fusion [correction of factor: a fusion] protein.
abstract: Locoregional and intratumoral administration of tumor necrosis factor alpha (TNF alpha) has been successful in obtaining inhibition or regression of tumor growth in the clinic. This potent antitumor activity of TNF alpha has not yet been exploited as a systemic agent in cancer therapy, mainly due to high levels of toxicity to normal tissues before a therapeutic dose of TNF alpha in the tumor has been achieved. To address this, we have targeted TNF alpha using antitumor antibodies. We have used a genetic fusion of human recombinant TNF alpha with MFE-23, a single-chain Fv antibody fragment directed against carcinoembryonic antigen. MFE-23::TNF alpha fusion protein is isolated in high yields (28 mg/L) from bacterial inclusion bodies and purified to homogeneity by affinity chromatography. It is a 144 kDa trimer in native form and possesses the antigen-binding activity of the sFv and the cytotoxicity to both WEHI 164 and a human adenocarcinoma cell line (LoVo) of rhTNF alpha. Radiolabeled MFE-23::TNF alpha binds both human and mouse TNF receptor 1 in vitro and is able to localize effectively in nude (nu/nu) mice bearing human LS174T xenografts; tumor/tissue ratios of 21:1 and 60:1 are achieved 24 and 48 h after intravenous injection. These studies indicate that MFE-23::TNF alpha will provide an effective means for systemically administered cancer therapy with TNF alpha.
authors.affiliation: CRC Targeting and Imaging Group, Academic Department of Oncology, Royal Free and University College Medical School, University College, London, NW3 2PF, UK. s.cooke@ucl.ac.uk
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Cooke
authors.forename: R Barbara
authors.initials: RB
authors.lastname: Pedley
authors.forename: R
authors.initials: R
authors.lastname: Boden
authors.forename: Richard H J
authors.initials: RH
authors.lastname: Begent
authors.forename: Kerry A
authors.initials: KA
authors.lastname: Chester
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000493:pharmacokinetics; D002272:Carcinoembryonic Antigen / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D016503:Drug Delivery Systems; D006801:Humans; D018796:Immunoconjugates / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D007128:Immunoglobulin Fragments / Q000008:administration & dosage* / Q000235:genetics / Q000494:pharmacology*; D051379:Mice; D008819:Mice, Nude; D009369:Neoplasms / Q000188:drug therapy / Q000276:immunology; D017510:Protein Folding; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics
chemical_list: D000970:Antineoplastic Agents; D002272:Carcinoembryonic Antigen; D018796:Immunoconjugates; D007128:Immunoglobulin Fragments; C451184:MFE-23 protein, human; D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; C055966:immunoglobulin Fv