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Sensors 2008, 8(8), 4669-4686; doi:10.3390/s8084669

Antibody Fragments as Probe in Biosensor Development

1,2,* , 1,2,3,4, 5 and 1,2
1 Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium 2 Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium 3 Department of Molecular Biology, Technologiepark 927, B-9052 Zwijnaarde, Ghent University, Ghent, Belgium 4 Department for Molecular Biomedical Research, VIB, Ghent, Belgium 5 IMEC, NEXT, Kapeldreef 75, B-3001 Leuven, Belgium
* Author to whom correspondence should be addressed.
Received: 7 July 2008 / Revised: 6 August 2008 / Accepted: 7 August 2008 / Published: 8 August 2008
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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.
Keywords: display technology; affinity; stability; immobilization; immunoassay. display technology; affinity; stability; immobilization; immunoassay.
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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Saerens, D.; Huang, L.; Bonroy, K.; Muyldermans, S. Antibody Fragments as Probe in Biosensor Development. Sensors 2008, 8, 4669-4686.

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