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Review
. 2008 Aug 8;8(8):4669-4686.
doi: 10.3390/s8084669.

Antibody Fragments as Probe in Biosensor Development

Dirk Saerens  1   2 , Lieven Huang  3   4   5   6 , Kristien Bonroy  7 , Serge Muyldermans  3   4
Affiliations
Review

Antibody Fragments as Probe in Biosensor Development

Dirk Saerens et al. Sensors (Basel). .

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.

Keywords: affinity; display technology; immobilization; immunoassay.; stability.

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Figures

Figure 1.
Figure 1.
Ab fragments from conventional (A), Heavy-chain (B) and cartilaginous fish (C).
Figure 2.
Figure 2.
Ab fragment immobilization via random (A) or oriented (B) covalent coupling.
Figure 3.
Figure 3.
Ab fragment immobilization via intermediate layer.
Figure 4.
Figure 4.
Ab fragment immobilization via biotin-(strept)avidin interaction.
Figure 5.
Figure 5.
Ab fragment immobilization via fusion partner.
Figure 6.
Figure 6.
Ab fragment immobilization via affinity tag.
Figure 7.
Figure 7.
Ab fragment immobilization via bispecific construct containing a Ab fragment specific for the surface.
See this image and copyright information in PMC

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