Human IgG antibody profiles differentiate between symptomatic patients with and without colorectal cancer

Abstract OBJECTIVE: Patients with cancer have antibodies against tumour antigens. Characterising the antibody repertoire may provide insights into aberrant cellular mechanisms in cancer development, ultimately leading to novel diagnostic or therapeutic targets. The aim of this study was to characterise the antibody profiles in patients whose symptoms warranted colonoscopy, to see if there was a difference in patients with and without colorectal cancer. METHODS: Patients were recruited from a colonoscopy clinic. Individual serum samples from 43 patients with colorectal cancer and 40 patients with no cancer on colonoscopy were profiled on a 37 830 clone recombinant human protein array. Antigen expression was evaluated by quantitative reverse transcription-PCR and by immunohistochemistry on tissue microarrays. RESULTS: Using a sex- and age-matched training set, 18 antigens associated with cancer and 4 associated with the absence of cancer (p\u3c0.05) were identified and confirmed. To investigate the mechanisms triggering antibody responses to these antigens, antigen expression was examined in normal colorectal mucosa and colorectal carcinoma of the same patients. The identified antigens showed cellular accumulation (p53), aberrant cellular expression (high mobility group B1 (HMGB1)) and overexpression (tripartite motif-containing 28 (TRIM28), p53, HMGB1, transcription factor 3 (TCF3), longevity assurance gene homologue 5 (LASS5) and zinc finger protein 346 (ZNF346)) in colorectal cancer tissue compared with normal colorectal mucosa. CONCLUSIONS: It is demonstrated for the first time that screening high-density protein arrays identifies unique antibody profiles that discriminate between symptomatic patients with and without colorectal cancer. The differential expression of identified antigens suggests their involvement in aberrant cellular mechanisms in cance

Comprehensive Modeling of Multimode Fiber Sensors for Refractive Index Measurement and Experimental Validation

We propose and develop a comprehensive model for estimating the refractive index (RI) response over three potential sensing zones in a multimode fiber. The model has been developed based on a combined ray optics, Gaussian beam, and wave optics analysis coupled to the consideration of the injected interrogating lightwave characteristics and validated experimentally through the realization of three sensors with different lengths of stripped cladding sections as the sensing region. The experimental results highly corroborate and validate the simulation output from the model for the three RI sensing zones. The sensors can be employed over a very wide dynamic RI range from 1.316 to over 1.608 at a wavelength of 1550 nm, with the best resolution of 2.2447 × 10−5 RI unit (RIU) obtained in Zone II for a 1-cm sensor length

Waveguide Fabrication In UV-Photocurable Sol–Gel Materials: Influence Of The Photoinitiating System

In this paper we identify and explain the different chemical interactions involved between a sol–gel matrix and photoinitiators used in the fabrication of optical waveguides. A well-established sol–gel matrix composed of 3-methacryloxypropyltrimethoxysilane, zirconium n-propoxide and methacrylic acid was developed, and two different photoinitiators (Irgacure® 819 and 1800) were added to the host matrix. Optical microscopy was used to characterise the structure of the waveguides as a function of the photoinitiator nature and concentration, and aging of the hybrid sol–gel material. It is clearly demonstrated that the width of the waveguides is strongly influenced by the sol aging. Furthermore, it is shown that degradation of photoinitiators occurs during the sol–gel process. Oxidation of the phosphonyl groups by the zirconium complex accounts for this results

Platform for enhanced detection efficiency in luminescence-based sensors

Luminescence-based biochip measurement platforms are employed in a wide range of biological applications, such as biomedical diagnostics. Based on an understanding of the anisotropic emission properties of luminescence emitters close to a dielectric interface, a simple strategy for producing a better than 25-fold enhancement of the detected luminescence is presented. This strategy is demonstrated for low cost polymer platforms compatible with mass-production

Surface plasmon resonance-enhanced light interaction in an integrated ormocomp nanowire

An integrated ormocomp nanowire coated with gold metal layer is proposed and its optical characteristics with the effect of surface plasmon resonance (SPR) are studied. The integrated rib-like nanowire has a trapezoidal shape with sidewall angles of 75°. It is coated with 50 nm gold layer to introduce the SPR and enhance the evanescent field in the sensing region located at the dielectric/metal interface. The possible field modes, the normalized power confinement, and the SPR peak position of the nanowire are studied over the wavelength and the metal thickness by using the full-vectorial H-field FEM in quasi-TM mode. The attenuation coefficient of the nanowire, the SPR peak wavelength, and the wavelength shift is experimentally extracted for three different cladding materials. The redshift of the supermode coupling between the dielectric mode and the anti-symmetric supermode is observed with the higher cladding-index and larger metal thickness. The improvement of the power confinement in the sensing region with the SPR effect is ten times (10×) better than a previous similar study

A comparison of mono and multivalent linkers and their effect on the colloidal stability of nanoparticle and immunoassays performance

When designing devices for biomedical diagnostics, increasing the signal to noise ratio is often critical for achieving clinically relevant sensitivity and limits of detection (LOD). In antibody-based assays, the measured signal can be amplified through the replacement of molecular fluorophores with doped nanoparticles (NP). However, the benefits of using NPs can only be realized if the NPs are coated efficiently with detection antibody, have good colloidal stability and the ratio of specific to non-specific binding (NSB) is high enough. The main focus of this paper is on the optimization of the bioconjugation protocol for antibody labeling of NPs leading to improved assay performance. Two types of linkers were used: monovalent linkers (glutaraldehyde; sulfo-SMCC; and sulfo-SIAB), and three generations of dendrimers endowed with multivalent carboxylic functionality. Overall, the NP-IgG conjugates prepared using multivalent linkers showed a significantly lower LOD and higher sensitivity than their homo- or hetero-functional counterparts. The multivalent dendrimers also improved NP stability and reduced aggregation. Moreover, the dendrimers showed a higher reactivity with biological material, a feature that could significantly reduce the cost of high-throughput biodiagnostics tests. © 2010 Elsevier B.V. All rights reserved

Improving the sensitivity of immunoassays with PEG-COOH-like film prepared by plasma-based technique

Herein we report on a preparation and performance of stable, hydrophilic and biocompatible polymeric material suitable for functionalization of disposable substrates used in biosensors. This new material features -COOH surface groups cross-linked with ethylene glycol molecules and was prepared in situ on disposable, plastic substrate by high-throughput and environmentally friendly technique called plasma-enhanced chemical vapor deposition (PECVD). The film is grafted to the plasma activated plastic by sequential deposition of tetraethylorthosilicate, forming a bonding layer, and mixed vapors of acrylic acid and diethyleneglycol dimethylether (AA/PEG) that provide the desired functional groups forming a sensing, contact layer. A superior performance of the AA/PEG coating as suitable material for substrates in biomedical devices was demonstrated in a model fluorescence linked immunosorbent assay. The results were compared with other commonly used surface materials prepared by wet chemistry methods. The unique characteristic of the AA/PEG film is that the immunoassay can be executed without the need for a blocking step, typically using albumins, without negative consequences on the bioassay results. In fact, the superior quality of the materials modified with AA/PEG film was highlighted by improving the sensitivity of an immunoassay by two orders of magnitude when compared with substrates prepared by standard surface chemistry methods. Copyright © 2011 Wiley Periodicals, Inc

Synthesis, stabilization, and functionalization of silver nanoplates for biosensor applications

Silver nanoplates (NPTs) were prepared by a seed-mediated growth method with different diameters (d = 25, 32, 53, and 100 nm) and with thicknesses of approximately 10 nm in all cases. As the concentration of silver seeds increased, the diameter of the nanoplates increased, resulting in an overall shift in the localized surface plasmon resonance (LSPR) band maximum from 570 to 900 nm, thus providing a novel method to tune the plasmon resonance. The LSPR was calculated from theory for both triangular and circular nanoplate geometries. In agreement with transmission electron micrographs, the model results confirmed the shape of nanoplates as being truncated prisms, intermediate between that of a prism and a disk. Because of the toxicity of the surfactant hexadecyltrimethylammonium bromide (CTAB), the stabilizing CTAB bilayer surrounding the NPT was replaced by a nontoxic alkanethiol with surfactant properties. This enabled the extraction of metal nanoparticles into deionized water or buffer for bioconjugation without aggregation. Silver nanoplates were also coated with polyelectrolyte layers using the standard layer-by-layer (LbL) method. The LSPR was found to be very sensitive to the addition of polyelectrolyte layers, with a plasmon band shift from 728 to 740 nm after adding only one monolayer (thickness ~1.5 nm). Bioconjugation of these nanoplates was achieved with the addition of a mercaptolinker containing a carboxyl group. The carboxyl groups were activated with 1-ethyl-3-(3- dimethylaminopropyl) hydrochloride (EDC)/N-hydroxysuccinimide (NHS) and conjugated to green fluorescent protein (GFP) in order to validate the potential of the NPTs for enhancement of bioassays. The fluorescence of the conjugated NPTs was 5.6-fold brighter than that of NPTs added to GFP without activation. © 2009 American Chemical Society

TIRF microscopy as a screening method for non-specific binding on surfaces

We report a method for studying nanoparticle-biosensor surface interactions based on total internal reflection fluorescence (TIRF) microscopy. We demonstrate that this simple technique allows for high throughput screening of non-specific adsorption (NSA) of nanoparticles on surfaces of different chemical composition. Binding events between fluorescent nanoparticles and functionalized Zeonor® surfaces are observed in real-time, giving a measure of the attractive or repulsive properties of the surface and the kinetics of the interaction. Three types of coatings have been studied: one containing a polymerized aminosilane network with terminal -NH2 groups, a second film with a high density of -COOH surface groups and the third with sterically restraining branched poly(ethylene)glycol (PEG) functionality. TIRF microscopy revealed that the NSA of nanoparticles with negative surface charge on such modified coatings decreased in the following order -NH2>-branched PEG>-COOH. The surface specificity of the technique also allows discrimination of the degree of NSA of the same surface at different pH. © 2010 Elsevier Inc