{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T13:39:53Z","timestamp":1777469993085,"version":"3.51.4"},"reference-count":31,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2002,2,25]],"date-time":"2002-02-25T00:00:00Z","timestamp":1014595200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Intl Journal of Cancer"],"published-print":{"date-parts":[[2002,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>A number of enzyme\/prodrug activation approaches for the treatment of cancer have been reported to date with varying success. We describe progress in the development of a system based on a \u03b2\u2010glucosidase enzyme in combination with a naturally occurring \u201cprodrug,\u201d the sugar linamarin, which releases the cytotoxin cyanide. A recombinant fusion protein, composed of an scFv (MFE\u201023) reactive against carcinoembryonic antigen (CEA) and a plant\u2010derived \u03b2\u2010glucosidase (linamarase), was produced and its cytotoxic potential was investigated. The fusion protein was expressed in a supersecretory mutant strain of <jats:italic>Saccharomyces cerevisiae<\/jats:italic> and purified by affinity chromatography. Extensive functional <jats:italic>in vitro<\/jats:italic> characterisation of the fusion protein showed that it retained antigen binding activity but that its catalytic activity was impaired, a problem not related to its fusion with the scFv. Nevertheless, we demonstrated complete tumour cell killing at doses of prodrug that are completely nontoxic to nontargeted cells. Preliminary <jats:italic>in vivo<\/jats:italic> characterisation showed that extensive glycosylation of the fusion protein caused its rapid clearance through the hepatic route. Aggregational properties also led to poor pharmacokinetics. Furthermore, we present some data analysing the mode of cell death resulting from exposure to this system. Enzymic catalysis of the substrate generates cyanide, a metabolic poison that asphyxiates cells and leads them to a necrotic\u2010like cell death. This system has been called antibody\u2010guided enzyme nitrile therapy (AGENT). \u00a9 2002 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/ijc.10266","type":"journal-article","created":{"date-parts":[[2002,8,25]],"date-time":"2002-08-25T23:16:32Z","timestamp":1030317392000},"page":"138-148","source":"Crossref","is-referenced-by-count":24,"title":["Antibody\u2010guided enzyme therapy of cancer producing cyanide results in necrosis of targeted cells"],"prefix":"10.1002","volume":"99","author":[{"given":"Christina A.","family":"Kousparou","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Agamemnon A.","family":"Epenetos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mahendra P.","family":"Deonarain","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2002,3,27]]},"reference":[{"key":"e_1_2_6_2_2","first-page":"55","article-title":"Advances in tumour targeting","volume":"1","author":"Kousparou CA","year":"2000","journal-title":"J Int Soc Tumour Targeting"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-5699(00)01668-6"},{"key":"e_1_2_6_4_2","first-page":"1711","article-title":"Radioimmunoguided surgery in colorectal cancer using a genetically engineered anti\u2010CEA single\u2010chain Fv antibody","volume":"6","author":"Mayer A","year":"2000","journal-title":"Clin Cancer Res"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/bjc.1987.237"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/bjc.1994.400"},{"key":"e_1_2_6_7_2","first-page":"5274","article-title":"The \u201cbystander effect\u201d: tumor regression when a fraction of the tumor mass is genetically modified","volume":"53","author":"Freeman SM","year":"1993","journal-title":"Cancer Res"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0215(20000215)85:4<571::AID-IJC20>3.0.CO;2-1"},{"key":"e_1_2_6_9_2","first-page":"765","article-title":"Antibody\u2010directed enzyme prodrug therapy: efficacy and mechanism of action in colorectal carcinoma","volume":"6","author":"Napier MP","year":"2000","journal-title":"Clin Cancer Res"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.25.15804"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-9861(92)90518-2"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0215(19981209)78:6<712::AID-IJC8>3.0.CO;2-D"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3300751"},{"key":"e_1_2_6_14_2","series-title":"Toxicology profile for cyanide","author":"Agency for"},{"key":"e_1_2_6_15_2","volume-title":"Molecular cloning: a laboratory manual","author":"Sambrook J","year":"1989"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1042\/0264-6021:3530199"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.3939723"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0215(20000515)86:4<561::AID-IJC19>3.0.CO;2-5"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0031-9422(00)86175-0"},{"key":"e_1_2_6_20_2","unstructured":"1998 Freeman New York A Fersht Structure and mechanism in protein science. 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