title: New strategies in polypeptide and antibody synthesis: an overview.
abstract: The synthesis of radioligands can benefit considerably from optimized recombinant protein production, both on the aspect of economy of production and on the level of improving the targeting and pharmacokinetics of the ligand. This paper first describes a general production optimization strategy, and then elaborates on a protein design strategy tailored to targeting applications. Production in Escherichia coli will benefit from economy of goods and time as compared to other organisms. In order to increase the chance of finding a successful production system in this host, we have assembled a large number of expression strategies in a single, uniform expression system (FastScreen). The system allows rapid optimization of direct production of native proteins or via a fusion protein strategy with subsequent recovery of the desired protein. As an example of recombinant radioligand synthesis for improved targeting and clearing, a manifold of intermediate molecular size was synthesized by fusing one Fab and two single-chain variable fragments (scFv) antibody binding fragments into a trifunctional molecule (Tribody). Due to the use of the specific heterodimerization of the Fab chains, trispecific, bispecific, or trivalent antibody derived targeting reagents can easily be obtained. Recombinant production techniques also allow for specific incorporation of amino acids favoring a site specific labeling (labeling tags).
authors.affiliation: Department of Molecular Biomedical Research, Flanders Interuniversity Institute of Biotechnology (VIB), Ghent University, Ghent, Belgium. Nico.Mertens@UGent.be
authors.forename: Nico
authors.initials: N
authors.lastname: Mertens
authors.forename: Filip
authors.initials: F
authors.lastname: Devos
authors.forename: Jannick
authors.initials: J
authors.lastname: Leoen
authors.forename: Els
authors.initials: E
authors.lastname: Van Deynse
authors.forename: An
authors.initials: A
authors.lastname: Willems
authors.forename: Steve
authors.initials: S
authors.lastname: Schoonooghe
authors.forename: Ingrid
authors.initials: I
authors.lastname: Burvenich
authors.forename: Stefaan
authors.initials: S
authors.lastname: De Koker
authors.forename: Dominique
authors.initials: D
authors.lastname: Vlieghe
authors.forename: Johan
authors.initials: J
authors.lastname: Grooten
authors.forename: Andrew
authors.initials: A
authors.lastname: Kelly
authors.forename: Christophe
authors.initials: C
authors.lastname: Van de Wiele
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D010455:Peptides; D011994:Recombinant Proteins