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Biosensors (Basel). 2016 Oct 11;6(4). pii: E52.

Kinetics of Antibody Binding to Membranes of Living Bacteria Measured by a Photonic Crystal-Based Biosensor.

Author information

1
Laboratoire de Physique de la Matière Vivante, IPHYS, École Polytechnique Fédérale de Lausanne (EPFL); Rte de la Sorge, 1015 Lausanne, Switzerland. ekaterina.rostova@epfl.ch.
2
Laboratoire de Physique de la Matière Vivante, IPHYS, École Polytechnique Fédérale de Lausanne (EPFL); Rte de la Sorge, 1015 Lausanne, Switzerland. carine.benadiba@epfl.ch.
3
Laboratoire de Physique de la Matière Vivante, IPHYS, École Polytechnique Fédérale de Lausanne (EPFL); Rte de la Sorge, 1015 Lausanne, Switzerland. giovanni.dietler@epfl.ch.
4
Laboratoire de Physique de la Matière Vivante, IPHYS, École Polytechnique Fédérale de Lausanne (EPFL); Rte de la Sorge, 1015 Lausanne, Switzerland. serguei.sekatski@epfl.ch.

Abstract

Optical biosensors based on photonic crystal surface waves (PC SWs) offer a possibility to study binding interactions with living cells, overcoming the limitation of rather small evanescent field penetration depth into a sample medium that is characteristic for typical optical biosensors. Besides this, simultaneous excitation of s- and p-polarized surface waves with different penetration depths is realized here, permitting unambiguous separation of surface and volume contributions to the measured signal. PC-based biosensors do not require a bulk signal correction, compared to widely used surface plasmon resonance-based devices. We developed a chitosan-based protocol of PC chip functionalization for bacterial attachment and performed experiments on antibody binding to living bacteria measured in real time by the PCSW-based biosensor. Data analysis reveals specific binding and gives the value of the dissociation constant for monoclonal antibodies (IgG2b) against bacterial lipopolysaccharides equal to KD = 6.2 ± 3.4 nM. To our knowledge, this is a first demonstration of antibody-binding kinetics to living bacteria by a label-free optical biosensor.

KEYWORDS:

antibody; bacteria; binding kinetics; label-free biosensor; ligand-receptor interaction; photonic crystal

PMID:
27727183
PMCID:
PMC5192372
DOI:
10.3390/bios6040052
[Indexed for MEDLINE]
Free PMC Article
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