Open AccessMolecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation
Author(s) -
Klaus Bosslet,
Jörg Czech,
Petra Lorenz,
H.H. Sedlacek,
Marcus Schuermann,
G. Seemann
Publication year - 1992
Publication title -
british journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.833
H-Index - 236
eISSN - 1532-1827
pISSN - 0007-0920
DOI - 10.1038/bjc.1992.47
Subject(s) - fusion protein , prodrug , in vivo , recombinant dna , avidity , enzyme , monoclonal antibody , immunoglobulin light chain , affinity chromatography , biological activity , microbiology and biotechnology , chemistry , immunoconjugate , biochemistry , in vitro , antibody , biology , immunology , gene
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.