SPR immunosensor for the detection of okadaic acid in mussels using magnetic particles as antibody carriers

Protein G-coated magnetic particles (MPs) were used as immobilisation supports for an antibody against okadaic acid (MAb(OA)) and carriers into a surface plasmon resonance (SPR) device for the development of a direct competitive immunosensor for okadaic acid (OA). SPR analysis of MAb(OA)-MP conjugates demonstrated that conjugations were successful with complete immobilisation of all the antibody biomolecules onto the MPs. Moreover, MAb(OA)-MP conjugates provided up to 11-fold higher SPR signals, compared to free MAb(OA). The use of conjugates in the direct competition assay provided a 3-fold lower LOD mu g/L (2.6 mu g of OA/L, equivalent to 12 mu g of OA/kg mussel meat). The presence of mussel matrix did not interfere in the OA quantification as seen in the calibration curves. Mussel samples, obtained from Ebro Delta's bays (NW Mediterranean) during a diarrheic shellfish poisoning (DSP) event and in the presence of Dinophysis sacculus, an OA producer, in the shellfish production area, were analysed with the MP-based SPR immunosensor. The OA contents correlated with those obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (y = 0.984x -5.273, R-2 = 0.789, p <0.001) and by mouse bioassay (MBA).

Production of a broad specificity antibody for the development and validation of an optical SPR screening method for free and intracellular microcystins and nodularin in cyanobacteria cultures

A highly sensitive broad specificity monoclonal antibody was produced and characterised for microcystin detection through the development of a rapid surface plasmon resonance (SPR) optical biosensor based immunoassay. The antibody displayed the following cross-reactivity: MC-LR 100%; MC-RR 108%; MC-YR 68%; MC-LA 69%; MC-LW 71%; MC-LF 68%; and Nodularin 94%. Microcystin-LR was covalently attached to a CM5 chip and with the monoclonal antibody was employed in a competitive 4min injection assay to detect total microcystins in water samples below the WHO recommended limit (1µg/L). A 'total microcystin' level was determined by measuring free and intracellular concentrations in cyanobacterial culture samples as this toxin is an endotoxin. Glass bead beating was used to lyse the cells as a rapid extraction procedure. This method was validated according to European Commission Decision 96/23/EC criteria. The method was proven to measure intracellular microcystin levels, the main source of the toxin, which often goes undetected by other analytical procedures and is advantageous in that it can be used for the monitoring of blooms to provide an early warning of toxicity. It was shown to be repeatable and reproducible, with recoveries from spiked samples ranging from 74 to 123%, and had % CVs below 10% for intra-assay analysis and 15% for inter-assay analysis. The detection capability of the assay was calculated as 0.5ng/mL for extracellular toxins and 0.05ng/mL for intracellular microcystins. A comparison of the SPR method with LC-MS/MS was achieved by testing six Microcystis aeruginosa cultures and this study yielded a correlation R(2) value of 0.9989.

Application du SPR dans le criblage des ligands synthétiques du CD36 et sa validation

Le CD36 est un récepteur de type éboueur de classe B exprimé à la surface de nombreux types cellulaires dont les macrophages, les cellules endothéliales de la microvasculature et les plaquettes. Ce récepteur multiligand est impliqué dans plusieurs processus pathologiques notamment l’athérosclérose, l’angiogénèse et la malaria via la liaison spécifique de ligands comme les lipoprotéines oxydées de basse densité, la thrombospondine-1 et la protéine PfEMP-1, respectivement. Les peptides de la relâche de l’hormone de croissance (GHRP) ont été identifiés comme les premiers ligands synthétiques du CD36. Afin de développer de nouveaux ligands synthétiques du CD36, l’établissement d’une méthode de criblage est essentiel pour découvrir des composés avec une liaison de haute affinité pour ce récepteur. Pour y parvenir, nous avons surexprimé le domaine extracellulaire du CD36 humain dans les cellules d’insectes Sf9. La protéine soluble purifiée par chromatographie d’affinité fut immobilisée à la surface d’une plaque de résonance de plasmons de surface (SPR) pour les études de liaison. La méthodologie développée a permis de caractériser les ligands du CD36 en déterminant leurs constantes de dissociation (KD), et d’établir une relation structure-activité des ligands de la famille des azapeptides, des composés dérivés du GHRP-6. Afin de valider la méthode par spectroscopie SPR, une corrélation a été établie entre les valeurs de KD obtenues en SPR et les valeurs d’CI50 de courbes d’inhibition de la phosphorylation des MAP kinases JNK1/2 induite par un phospholipide oxydé, le POVPC, en présence de concentrations croissantes de ligands du CD36 dans les macrophages RAW 264.7.

I kolerans spår : En studie om koleraepidemins utveckling i Alingsås med omnejd 1834

Syftet med denna uppsats är att dels visa koleraepidemins utbredning i Alingsås år 1834 samt att kort jämföra den med omgivande städer i Västsverige och då framförallt Borås, och dels se på vilka återverkningar, upplevelser och reaktioner sjukdomen skapade hos människorna vid denna tid. De frågeställningar som har använts för att utveckla ovanstående syfte har varit: Vilka tankar, intryck och upplevelser gav sjukdomen upphov till? Vilka drabbades hårdast av sjukdomen? Hur förhåller sig dödstal, ålder, tidsdatering och social tillhörighet bland döda i en jämförelse med Borås? Vilka rykten, myter och reaktioner följde i kolerans spår? Materialet består av dödböcker och husförhörslängder för Alingsås stad och landsförsamling, protokoll från sundhetsnämnden i Alingsås landsförsamling, kungörelser och nedskrivna samtal från 1834, dagbok och anteckningar från privatpersoner från nämnda år samt litteratur och avhandlingar om Alingsås och koleran i Sverige. Alingsås stad hade vid sjukdomens inledningsskede en viss brist på läkare men detta avhjälptes snabbt och situationen var inte värre än i andra städer vid denna tid. För landsförsamlingen var dock bristen betydande. Då koleran härjade som värst upplevde människorna i Alingsås, liksom i många andra drabbade städer, en stark oro och ångest. De som oftast föll offer för koleran i Alingsås var fattiga som troligtvis på grund av trångboddhet och hygieniska svårigheter hade svårt att hålla smittan ifrån sig. I jämförelse med Borås visar Alingsås upp många likheter men också skillnader i form av fler döda och större dödlighet bland de fattiga.

Utveckling av metod för att bedöma behovet av ogräsbekämpning i spår

A system for weed management on railway embankments that is both adapted to the environment and efficient in terms of resources requires knowledge and understanding about the growing conditions of vegetation so that methods to control its growth can be adapted accordingly. Automated records could complement present-day manual inspections and over time come to replace these. One challenge is to devise a method that will result in a reasonable breakdown of gathered information that can be managed rationally by affected parties and, at the same time, serve as a basis for decisions with sufficient precision. The project examined two automated methods that may be useful for the Swedish Transport Administration in the future: 1) A machine vision method, which makes use of camera sensors as a way of sensing the environment in the visible and near infrared spectrum; and 2) An N-Sensor method, which transmits light within an area that is reflected by the chlorophyll in the plants. The amount of chlorophyll provides a value that can be correlated with the biomass. The choice of technique depends on how the information is to be used. If the purpose is to form a general picture of the growth of vegetation on railway embankments as a way to plan for maintenance measures, then the N-Sensor technique may be the right choice. If the plan is to form a general picture as well as monitor and survey current and exact vegetation status on the surface over time as a way to fight specific vegetation with the correct means, then the machine vision method is the better of the two. Both techniques involve registering data using GPS positioning. In the future, it will be possible to store this information in databases that are directly accessible to stakeholders online during or in conjunction with measures to deal with the vegetation. The two techniques were compared with manual (visual) estimations as to the levels of vegetation growth. The observers (raters) visual estimation of weed coverage (%) differed statistically from person to person. In terms of estimating the frequency (number) of woody plants (trees and bushes) in the test areas, the observers were generally in agreement. The same person is often consistent in his or her estimation: it is the comparison with the estimations of others that can lead to misleading results. The system for using the information about vegetation growth requires development. The threshold for the amount of weeds that can be tolerated in different track types is an important component in such a system. The classification system must be capable of dealing with the demands placed on it so as to ensure the quality of the track and other pre-conditions such as traffic levels, conditions pertaining to track location, and the characteristics of the vegetation. The project recommends that the Swedish Transport Administration: Discusses how threshold values for the growth of vegetation on railway embankments can be determined Carries out registration of the growth of vegetation over longer and a larger number of railway sections using one or more of the methods studied in the project Introduces a system that effectively matches the information about vegetation to its position Includes information about the growth of vegetation in the records that are currently maintained of the track’s technical quality, and link the data material to other maintenance-related databases Establishes a number of representative surfaces in which weed inventories (by measuring) are regularly conducted, as a means of developing an overview of the long-term development that can serve as a basis for more precise prognoses in terms of vegetation growth Ensures that necessary opportunities for education are put in place

A S-parameters-based detection method for a multilayer SPR biosensor

In this paper, S-parameters investigation of a variable incidence angle multilayer SPR biosensor is presented. Both magnitude and phase of the S-parameters are taken into account in the investigation. The work presented in this paper is the first attempt to apply S-parameters analysis to a multilayer SPR biosensor. The goal is to improve sensitivity through involving S-parameters including their phase values. In addition, further investigation is carried out to understand the relationship between the S-parameters and thickness of biomolecular layer and also the design parameters including the number of graphene layers.

Variable incidence angle subwavelength grating SPR graphene biosensor

This paper investigates the sensitivity enhancement of a variable incidence angle subwavelength grating based multilayer surface plasmon resonance biosensor (SPRB). In the proposed design, a periodic array of subwavelength grating is integrated on top of a layer of graphene sheet in the multilayer SPR biosensor. The performance of the biosensor is investigated through monitoring the biomolecular interactions of cDNA-ssDNA interactions on its surface. The sensitivity improvement is indicated by the shift of the resonance peak angle.

Investigação numérica do desempenho de um sensor de fibra ótica de índice de refração, baseado em SPR e utilizando COMSOL multiphysics

Esta tese é dedicada aos sensores de fibra ótica especificamente aos sensores baseados no fenómeno de ressonância de plasmão de superfície, SPR (Surface Plasmon Resonance), gerados em fibras óticas com configuração do tipo “D”, para aplicação em sensores refratométricos. Numa primeira parte desta dissertação são descritos os aspetos teóricos fundamentais para a compreensão dos fenómenos de ressonância de plasmões de superfície e a sua utilização em sistemas sensores. Estes fenómenos ocorrem na superfície de interface entre metais e outros meios materiais, sendo capazes de afetar as propriedades em reflexão e transmissão de uma onda eletromagnética incidente (feixe luminoso), de uma forma que é fortemente dependente dos meios na proximidade do metal. Assim, a medição das propriedades do feixe luminoso, como por exemplo o comprimento de onda de ressonância com SPR, permite monitorizar esses meios. Numa segunda fase foi implementada a simulação destes modelos, em COMSOL Multiphysics, que permitia não só a obtenção dos espetros de transmissão dos fenómenos de ressonância de plasmões de superfície, mas também a obtenção das distribuições do campo elétrico e magnético em função das dimensões do sensor. O COMSOL permitiu também a obtenção das curvas do deslocamento do comprimento de onda ressonante, perante variações do índice de refração exterior, da espessura do metal, da espessura da bainha e da espessura de outro elemento de elevado índice de refração. A fase seguinte foi verificar que os resultados dos métodos teóricos para os diferentes parâmetros de estudo eram semelhantes aos resultados obtidos no COMSOL. Conclui-se que com este programa é possível criar novos sensores em fibra ótica, baseados em SPR, para melhorar e otimizar os parâmetros de ressonância e sensibilidade do sensor. A última fase do trabalho baseou-se na modelização de uma fibra cuja configuração seja tal que possa criar um pequeno efeito de antena e fazer com que parte da luz seja guiada para o exterior da fibra e possa interatuar com o meio externo para melhor sensibilidade.

Design of SPR systems with dynamic compensators and output variable structure control

This paper presents necessary and sufficient conditions for the following problem: given a linear time invariant plant G(s) = N(s)D(s)-1 = C(sI - A]-1B, with m inputs, p outputs, p > m, rank(C) = p, rank(B) = rank(CB) = m, £nd a tandem dynamic controller Gc(s) = D c(s)-1Nc(s) = Cc(sI - A c)-1Bc + Dc, with p inputs and m outputs and a constant output feedback matrix Ko ε ℝm×p such that the feedback system is Strictly Positive Real (SPR). It is shown that this problem has solution if and only if all transmission zeros of the plant have negative real parts. When there exists solution, the proposed method firstly obtains Gc(s) in order to all transmission zeros of Gc(s)G(s) present negative real parts and then Ko is found as the solution of some Linear Matrix Inequalities (LMIs). Then, taking into account this result, a new LMI based design for output Variable Structure Control (VSC) of uncertain dynamic plants is presented. The method can consider the following design specifications: matched disturbances or nonlinearities of the plant, output constraints, decay rate and matched and nonmatched plant uncertainties. © 2006 IEEE.

Variable structure control of switched systems based on Lyapunov-Metzler-SPR systems

This paper presents two Variable Structure Controllers (VSC) for continuous-time switched plants. It is assumed that the state vector is available for feedback. The proposed control system provides a switching rule and also the variable structure control input. The design is based on Lyapunov-Metzler (LM) inequalities and also on Strictly Positive Real (SPR) systems stability results. The definition of Lyapunov-Metzler-SPR (LMS) systems and its direct application in the design of VSC for switched systems are introduced in this paper. Two examples illustrate the design of the proposed VSC, considering a plant given by a switched system with a switched-state control law and two linear time-invariant systems, that are not controllable and also can not be stabilized with state feedback. ©2008 IEEE.

Comparative study of LMI-based output feedback spr synthesis for plants with different numbers of inputs and outputs

New Linear Matrix Inequalities (LMI) conditions are proposed for the following problem, called Strictly Positive Real (SPR) synthesis: given a linear time-invariant plant, find a constant output feedback matrix Ko and a constant output tandem matrix F for the controlled system to be SPR. It is assumed that the plant has the number of outputs greater than the number of inputs. Some sufficient conditions for the solution of the problem are presented and compared. These results can be directly applied in the LMI-based design of Variable Structure Control (VSC) of uncertain plants. ©2008 IEEE.

A immunosensor for the diagnosis of canine distemper virus infection using SPR and EIS

Canine distemper virus (CDV) is a viral disease that affects dogs and many other carnivores. Clinical diagnosis of CDV is difficult due to the broad spectrum of signs that may be confounded with other respiratory and enteric diseases of dogs. Laboratory analysis is required to diagnose suspected cases. In this study, surface plasmon resonance (SPR) and electrochemical impedance spectroscopy (EIS) methodologies were developed for the detection of canine distemper virus simultaneously. The assay exhibited high specificity, as all the negative controls were not mistakenly detected. The CDV concentration was determined from successive injections into the apparatus, with a linear range from 1.1 to 116.0 ng mL-1. The system exhibited good reproducibility with 4.5% variation between runs after regeneration of the coated surface with a solution of 0.1 M glycine-HCL (pH 3.0). The capacitance and resistance values of the modified interface were calculated from EIS data using an equivalent circuit. It was possible to measure CDV in highly concentrated viruses with good specificity and reproducibility. © 2013 The Royal Society of Chemistry.