title: Antibody Fragments as Probe in Biosensor Development.
abstract: Today's proteomic analyses are generating increasing numbers of biomarkers, making it essential to possess highly specific probes able to recognize those targets. Antibodies are considered to be the first choice as molecular recognition units due to their target specificity and affinity, which make them excellent probes in biosensor development. However several problems such as difficult directional immobilization, unstable behavior, loss of specificity and steric hindrance, may arise from using these large molecules. Luckily, protein engineering techniques offer designed antibody formats suitable for biomarker analysis. Minimization strategies of antibodies into Fab fragments, scFv or even single-domain antibody fragments like VH, VL or VHHs are reviewed. Not only the size of the probe but also other issues like choice of immobilization tag, type of solid support and probe stability are of critical importance in assay development for biosensing. In this respect, multiple approaches to specifically orient and couple antibody fragments in a generic one-step procedure directly on a biosensor substrate are discussed.
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium. dsaerens@vub.ac.be.|Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium. dsaerens@vub.ac.be.
authors.forename: Dirk
authors.initials: D
authors.lastname: Saerens
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium.|Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium.|Department of Molecular Biology, Technologiepark 927, B-9052 Zwijnaarde, Ghent University, Ghent, Belgium.|Department for Molecular Biomedical Research, VIB, Ghent, Belgium.
authors.forename: Lieven
authors.initials: L
authors.lastname: Huang
authors.affiliation: IMEC, NEXT, Kapeldreef 75, B-3001 Leuven, Belgium.
authors.forename: Kristien
authors.initials: K
authors.lastname: Bonroy
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium.|Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium.
authors.forename: Serge
authors.initials: S
authors.lastname: Muyldermans
keywords: affinity; display technology; immobilization; immunoassay.; stability