title: Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.
abstract: A fusion protein consisting of the humanised Fab fragment of the anti CEA MAb BW 431 and the human beta-glucuronidase was expressed in BHK cells. Functional testing revealed that the specificity and avidity of the humanised V region was similar to the original murine MAb BW 431. Furthermore, the enzymatic activity, pH sensitivity and stability of the human beta-glucuronidase in the fusion protein was comparable to the activity of recombinant human beta-glucuronidase. Using anti-idiotype affinity chromatography, two molecules of a molecular weight of 125 kDa or 250 kDa could be visualized under nonreducing conditions in SDS-PAGE. Reducing conditions revealed a 25 kDa light and 100 kDa heavy chain. Due to its suitable biological characteristics this fusion protein might be an appropriate molecule allowing a site specific antibody directed enzyme prodrug therapy (ADEPT) in vivo.
authors.affiliation: Research Laboratory of Behringwerke, Marburg, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Bosslet
authors.forename: J
authors.initials: J
authors.lastname: Czech
authors.forename: P
authors.initials: P
authors.lastname: Lorenz
authors.forename: H H
authors.initials: HH
authors.lastname: Sedlacek
authors.forename: M
authors.initials: M
authors.lastname: Schuermann
authors.forename: G
authors.initials: G
authors.lastname: Seemann
mesh_terms: D002272:Carcinoembryonic Antigen / Q000378:metabolism; D002460:Cell Line; D005818:Genetic Engineering / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000235:genetics; D011355:Prodrugs / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism*; D014162:Transfection
chemical_list: D002272:Carcinoembryonic Antigen; D007074:Immunoglobulin G; D011355:Prodrugs; D011993:Recombinant Fusion Proteins