Clotilde Ribaut

Small biomolecule immunosensing with plasmonic optical fiber grating sensor

This study reports on the development of a surface plasmon resonance (SPR) optical fiber biosensor based on tilted fiber Bragg grating technology for direct detection of small biomarkers of interest for lung cancer diagnosis. Since SPR principle relies on the refractive index modifications to sensitively detect mass changes at the gold coated surface, we have proposed here a comparative study in relation to the target size. Two cytokeratin 7 (CK7) samples with a molecular weight ranging from 78 kDa to 2.6 kDa, respectively CK7 full protein and CK7 peptide, have been used for label-free monitoring. This work has first consisted in the elaboration and the characterization of a robust and reproducible bioreceptor, based on antibody/antigen cross-linking. Immobilized antibodies were then utilized as binding agents to investigate the sensitivity of the biosensor towards the two CK7 antigens. Results have highlighted a very good sensitivity of the biosensor response for both samples diluted in phosphate buffer with a higher limit of detection for the larger CK7 full protein. The most groundbreaking nature of this study relies on the detection of small biomolecule CK7 peptides in buffer and in the presence of complex media such as serum, achieving a limit of detection of 0.4 nM.

Fiber-Optic SPR Immunosensors Tailored To Target Epithelial Cells through Membrane Receptors

We report, for the first time, the use of a surface plasmon resonance (SPR) fiber-optic immunosensor for selective cellular detection through membrane protein targeting. The sensor architecture lies on gold-coated tilted fiber Bragg gratings (Au-coated TFBGs) photoimprinted in the fiber core via a laser technique. TFBGs operate in the near-infrared wavelength range at ∼1550 nm, yielding optical and SPR sensing characteristics that are advantageous for the analyses of cellular bindings and technical compatibility with relatively low-cost telecommunication-grade measurement devices. In this work, we take consider their numerous assets to figure out their ability to selectively detect intact epithelial cells as analytes in cell suspensions in the range of 2-5 × 10(6) cells mL(-1). For this, the probe was first thermally annealed to ensure a strong adhesion of the metallic coating to the fiber surface. Its surface was then functionalized with specific monoclonal antibodies via alkanethiol self-assembled monolayers (SAMs) against extracellular domain of epidermal growth factor receptors (EGFRs) and characterized by peak force tapping atomic force microscopy. A differential diagnosis has been demonstrated between two model systems. The developed immunosensors were able to monitor, in real time, the specific attachment of single intact cells in concentrations from 3 × 10(6) cells mL(-1). Such results confirm that the developed probe fits the lab-on-fiber technology and has the potential to be used as a disposable device for in situ and real-time clinical diagnosis.

Cancer biomarker sensing using packaged plasmonic optical fiber gratings: Towards in vivo diagnosis

This work presents the development of an innovative plasmonic optical fiber (OF) immunosensor for the detection of cytokeratin 17 (CK17), a biomarker of interest for lung cancer diagnosis. The development of this sensing platform is such that it can be assessed in non-liquid environments, demonstrating that a surface plasmon resonance (SPR) can be excited in this case. For this purpose, detections have been first carried out on CK17 encapsulated in gel matrix in the aim of mimicking tissue samples. Gold-coated OF immunosensors were embedded in a specifically designed packaging providing enough stiffness to penetrate into soft matters. Resulting reflected spectra have revealed, for the first time, the presence of a stable SPR signal recorded in soft matters. Experiments conducted to detect CK17 trapped in a porous polyacrylamide gel matrix have highlighted the specific and selective biosensor response towards the target protein. Finally, the packaged OF immunosensor has been validated by a preliminary test on human lung biopsy, which has confirmed the ex-vivo CK17 detection. Consequently, this work represents an important milestone towards the detection of biomarkers in tissues, which is still a clinical challenge for minimally-invasive in vivo medical diagnosis.

Immunosensing with Near-Infrared Plasmonic Optical Fiber Gratings

Surface Plasmon resonance (SPR) optical fiber biosensors constitute a miniaturized counterpart to the bulky prism configuration and offer remote operation in very small volumes of analyte. They are a cost-effective and relatively straightforward technique to yield in situ (or even possibly in vivo) molecular detection. They are usually obtained from a gold-coated fiber segment for which the core-guided light is brought into contact with the surrounding medium, either by etching (or side-polishing) or by using grating coupling. Recently, SPR generation was achieved in gold-coated tilted fiber Bragg gratings (TFBGs). These sensors probe the surrounding medium with near-infrared narrowband resonances, which enhances both the penetration depth of the evanescent field in the external medium and the wavelength resolution of the interrogation. They constitute the unique configuration able to probe all the fiber cladding modes individually, with high Q-factors. We use these unique spectral features in our work to sense proteins and extra-cellular membrane receptors that are both overexpressed in cancerous tissues. Impressive limit of detection (LOD) and sensitivity are reported, which paves the way for the further use of such immunosensors for cancer diagnosis.

Near-infrared spectral combs probe molecular interactions

Surface Plasmon resonance (SPR) optical fibre biosensors appear as a cost-effective and relatively straightforward technique to yield in situ (or even possibly in vivo) macromolecular detection. The biosensor configuration reported in this work uses nanometric-scale gold-coated tilted fibre Bragg gratings (TFBGs) interrogated by light polarized radially to the optical fibre outer surface. Recently, we have reported unprecedented surrounding refractive index (SRI) sensitivity for these SPR sensors, namely 10-5 refractive index unit. We report in this work the detection and quantification of cytokeratins in serum, which are proteins of interest for the lung cancer diagnosis. Results are presented in terms of sensitivity and limit of detection.

Visible vs near-infrared optical fiber plasmonics: performance comparison for protein sensing

In this work, two plasmonic optical fiber sensor configurations are used for protein sensing and their relative performances in terms of limit of detection and sensitivity are compared. The first configuration consists in unclad 200 μm optical fibers that produce a broadband resonance in the visible wavelength range around 650 nm while the second configuration makes use of multiple narrowband resonances produced in the C+L bands with weakly tilted fiber Bragg gratings photo-inscribed in telecommunication-grade single-mode optical fibers. In both cases, the sensitive regions are surrounded by a ~50 nm gold layer so that the evanescent wave can excite a surface plasmon polariton at the metalsurrounding medium interface. Both configurations are used to sense green fluorescent proteins. Our experimental results demonstrate that the two sensor configurations present a complementary measurement dynamics as a function of the investigated concentration in the range 10-12 – 10-7 g/ml. We attribute this difference of sensitivity to the difference of penetration depth of the evanescent wave in the surrounding medium, which is proportional to the light wavelength.