Identification of nonlinear lateral flow immunoassay state-space models via particle filter approach

This is the post-print of the Article. The official published version can be accessed from the link below - Copyright @ 2012 IEEEIn this paper, the particle filtering approach is used, together with the kernel smoothing method, to identify the state-space model for the lateral flow immunoassay through available but short time-series measurement. The lateral flow immunoassay model is viewed as a nonlinear dynamic stochastic model consisting of the equations for the biochemical reaction system as well as the measurement output. The renowned extended Kalman filter is chosen as the importance density of the particle filter for the purpose of modeling the nonlinear lateral flow immunoassay. By using the developed particle filter, both the states and parameters of the nonlinear state-space model can be identified simultaneously. The identified model is of fundamental significance for the development of lateral flow immunoassay quantification. It is shown that the proposed particle filtering approach works well for modeling the lateral flow immunoassay.This work was supported in part by the International Science and Technology Cooperation Project of China under Grant 2009DFA32050, Natural Science Foundation of China under Grants 61104041, International Science and Technology Cooperation Project of Fujian Province of China under Grant 2009I0016

Modulating Linker Composition of Haptens Resulted in Improved Immunoassay for Histamine.

Histamine (HA) is an important food contaminant generated during food fermentation or spoilage. However, an immunoassay for direct (derivatization free) determination of HA has rarely been reported due to its small size to induce the desired antibodies by its current hapten-protein conjugates. In this work, despite violating the classical hapten design criteria which recommend introducing a linear aliphatic (phenyl free) linker into the immunizing hapten, a novel haptens, HA-245 designed and synthesized with a phenyl-contained linker, exhibited significantly enhanced immunological properties. Thus, a quality-improved monoclonal antibody (Mab) against HA was elicited by its hapten-carrier conjugates. Then, as the linear aliphatic linker contained haptens, Hapten B was used as linker-heterologous coating haptens to eliminate the recognition of linker antibodies. Indirect competitive ELISA (ic-ELISA) was developed with a 50% inhibition concentration (IC50) of 0.21 mg/L and a limit of detection (LOD) of 0.06 mg/L in buffer solution. The average recoveries of HA from spiked food samples for this ic-ELISA ranged from 84.1% and 108.5%, and the analysis results agreed well with those of referenced LC-MS/MS. This investigation not only realized derivatization-free immunoassay for HA, but also provided a valuable guidance for hapten design and development of immunoassay for small molecules

Inference of nonlinear state-space models for sandwich-type lateral flow immunoassay using extended Kalman filtering

Copyright [2011] IEEE. This material is posted here with permission of the IEEE. Such permission of the IEEE does not in any way imply IEEE endorsement of any of Brunel University's products or services. Internal or personal use of this material is permitted. However, permission to reprint/republish this material for advertising or promotional purposes or for creating new collective works for resale or redistribution must be obtained from the IEEE by writing to [email protected]. By choosing to view this document, you agree to all provisions of the copyright laws protecting it.In this paper, a mathematical model for sandwichtype lateral flow immunoassay is developed via short available time series. A nonlinear dynamic stochastic model is considered that consists of the biochemical reaction system equations and the observation equation. After specifying the model structure, we apply the extend Kalman filter (EKF) algorithm for identifying both the states and parameters of the nonlinear state-space model. It is shown that the EKF algorithm can accurately identify the parameters and also predict the system states in the nonlinear dynamic stochastic model through an iterative procedure by using a small number of observations. The identified mathematical model provides a powerful tool for testing the system hypotheses and also inspecting the effects from various design parameters in a both rapid and inexpensive way. Furthermore, by means of the established model, the dynamic changes of the concentration of antigens and antibodies can be predicted, thereby making it possible for us to analyze, optimize and design the properties of lateral flow immunoassay devices.This work was supported in part by the International Science and Technology Cooperation Project of China under Grant 2009DFA32050, Natural Science Foundation of Fujian Province of China under Grants 2009J01280 and 2009J01281

The effects of zinc sulfate on ethyl glucuronide immunoassay urine testing

Published research in the Journal of Analytical Toxicology and the American Society for Clinical Pathology has confirmed that the presence of Zinc Sulfate in adulterated urine samples can influence the testing results using EMIT and ELISA immunoassay testing when testing for Cannabinoids (THC), Cocaine (Benzoylecgonine), Methamphetamines, Opiates (Morphine, Methadone, and Propoxyphene), Phencyclidine (PCP), and Ethanol (Alcohol Dehydrogenase). This research included adding Zinc Sulfate directly to urine samples. In 2006, the Substance Abuse and Mental Health Service Administration (SAMHSA) released an advisory that the use of Ethyl glucuronide (EtG) as a new biomarker as an indicator for the past-use of alcohol was promising and warranted more research. Ethyl glucuronide is a direct metabolite of the biotransformation of ethanol in the human body. This compound is excreted in urine and can be used as a specific biomarker for the ingestion of alcohol. Because EtG is only produced when ethanol is metabolized, there are no false positives due to fermentation and a much longer detection window exists for its detection. Scientific literature states that EtG can be present in urine long after ethanol has been eliminated. Testing for EtG is commonly referred to as the “80 hour test” for the ability of EtG to be measured up to 80 hours after consuming alcohol. It was hypothesized that if the presence of Zinc Sulfate added to urine falsely reduced urine alcohol level when measuring for Alcohol Dehydrogenase enzyme, will the presence of Zinc Sulfate added to SurineTM falsely reduce the urine alcohol level when measuring for EtG? Since it is very likely that EtG would still be present in the body after ethanol has been eliminated, samples contained either no ethanol or 5% (5g/dL) of ethanol. Samples were spiked at 10mg/mL, 15mg/mL or contained 0mg/mL of Zinc Sulfate. Additionally, duration testing was conducted to see if there was any observed differences between testing the samples fresh and then after a one week duration in a refrigerator and brought to room temperature prior to testing. Two different immunoassay EtG tests were used to perform the analysis. It was concluded that Zinc Sulfate directly added to the sample affected one of the immunoassay test regardless of whether EtG or ethanol were present, by fading the Test and Control regions. Additionally, it is concluded that SurineTM samples containing Zinc Sulfate could easily be distinguished from samples free of Zinc Sulfate because of the presence of a white cloudy precipitate

Multiplexed in-gel microfluidic immunoassays: characterizing protein target loss during reprobing of benzophenone-modified hydrogels.

From whole tissues to single-cell lysate, heterogeneous immunoassays are widely utilized for analysis of protein targets in complex biospecimens. Recently, benzophenone-functionalized hydrogel scaffolds have been used to immobilize target protein for immunoassay detection with fluorescent antibody probes. In benzophenone-functionalized hydrogels, multiplex target detection occurs via serial rounds of chemical stripping (incubation with sodium-dodecyl-sulfate (SDS) and β-mercaptoethanol at 50-60 °C for ≥1 h), followed by reprobing (interrogation with additional antibody probes). Although benzophenone facilitates covalent immobilization of proteins to the hydrogel, we observe 50% immunoassay signal loss of immobilized protein targets during stripping rounds. Here, we identify and characterize signal loss mechanisms during stripping and reprobing. We posit that loss of immobilized target is responsible for ≥50% of immunoassay signal loss, and that target loss is attributable to disruption of protein immobilization by denaturing detergents (SDS) and incubation at elevated temperatures. Furthermore, our study suggests that protein losses under non-denaturing conditions are more sensitive to protein structure (i.e., hydrodynamic radius), than to molecular mass (size). We formulate design guidance for multiplexed in-gel immunoassays, including that low-abundance proteins be immunoprobed first, even when targets are covalently immobilized to the gel. We also recommend careful scrutiny of the order of proteins targets detected via multiple immunoprobing cycles, based on the protein immobilization buffer composition

Three Dimensional Quantitative Structure-Activity Relationships of Sulfonamides Binding Monoclonal Antibody by Comparative Molecular Field Analysis

The three-dimensional quantitative structure-activity relationship (3D-QSAR) model of sulfonamide analogs binding a monoclonal antibody (MabSMR) produced against sulfamerazine, was carried out by comparative molecular field analysis (CoMFA). The affinities of MabSMR, expressed as Log10IC50, for 17 sulfonamide analogs were determined by competitive fluorescence polarization immunoassay (FPIA). Removal of two outliers from the initial set of 17 sulfonamide analogs improved the predictability of the models. The 3D-QSAR model of 15 sulfonamides resulted in q2cv values of 0.600, and r2 values of 0.995, respectively. This novel study combining FPIA with CoMFA demonstrates that multidisciplinary research can be used as a useful tool to investigate antigen-antibody interactions and provide information required for design of novel haptens, which may result in new antibodies with properties already optimized by an antibody-based immunoassay

A competitive enzyme immunoassay for the quantitative detection of cocaine from banknotes and latent fingermarks

A sensitive and versatile competitive enzyme immunoassay (cEIA) has been developed for the quantitative detection of cocaine in complex forensic samples. Polyclonal anti-cocaine antibody was purified from serum and deposited onto microtiter plates. The concentration of the cocaine antibody adsorbed onto the plates, and the dilution of the cocaine-HRP hapten were both studied to achieve an optimised immunoassay. The method was successfully used to quantify cocaine in extracts taken from both paper currency and latent fingermarks. The limit of detection (LOD) of 0.162 ng mL-1 achieved with the assay compares favourably to that of conventional chromatography-mass spectroscopy techniques, with an appropriate sensitivity for the quantification of cocaine at the low concentrations present in some forensic samples. The cEIA was directly compared to LC-MS for the analysis of ten UK banknote samples. The results obtained from both techniques were statistically similar, suggesting that the immunoassay was unaffected by cross-reactivity with potentially interfering compounds. The cEIA was used also for the detection of cocaine in extracts from latent fingermarks. The results obtained were compared to the cocaine concentrations detected in oral fluid sampled from the same individual. Using the cEIA, we have shown, for the first time, that endogeneously excreted cocaine can be detected and quantified from a single latent fingermark. Additionally, it has been shown that the presence of cocaine, at similar concentrations, in more than one latent fingermark from the same individual can be linked with those concentrations found in oral fluid. These results show that detection of drugs in latent fingermarks could directly indicate whether an individual has consumed the drug. The specificity and feasibility of measuring low concentrations of cocaine in complex forensic samples demonstrates the effectiveness and robustness of the assay. The immunoassay presents a simple and cost-effective alternative to the current mass spectrometry based techniques for the quantitation of cocaine at forensically significant concentrations

A hybrid EKF and switching PSO algorithm for joint state and parameter estimation of lateral flow immunoassay models

This is the post-print version of the Article. The official published can be accessed from the link below - Copyright @ 2012 IEEEIn this paper, a hybrid extended Kalman filter (EKF) and switching particle swarm optimization (SPSO) algorithm is proposed for jointly estimating both the parameters and states of the lateral flow immunoassay model through available short time-series measurement. Our proposed method generalizes the well-known EKF algorithm by imposing physical constraints on the system states. Note that the state constraints are encountered very often in practice that give rise to considerable difficulties in system analysis and design. The main purpose of this paper is to handle the dynamic modeling problem with state constraints by combining the extended Kalman filtering and constrained optimization algorithms via the maximization probability method. More specifically, a recently developed SPSO algorithm is used to cope with the constrained optimization problem by converting it into an unconstrained optimization one through adding a penalty term to the objective function. The proposed algorithm is then employed to simultaneously identify the parameters and states of a lateral flow immunoassay model. It is shown that the proposed algorithm gives much improved performance over the traditional EKF method.This work was supported in part by the International Science and Technology Cooperation Project of China under Grant 2009DFA32050, Natural Science Foundation of China under Grants 61104041, International Science and Technology Cooperation Project of Fujian Province of China under Grant 2009I0016

Graphene oxide-Au nano particle coated quartz crystal microbalance biosensor for the real time analysis of carcinoembryonic antigen

A label-free quartz crystal microbalance (QCM) biosensor was developed for the selective and real-time estimation of carcinoembryonic antigen (CEA) through the present study. Graphene oxide-Au nanoparticles (GO-AuNPs) was in situ synthesised on the surface of the QCM electrode and the antibody of CEA (monoclonal anti-CEA from mouse) was covalently immobilized on this layer as the bioreceptor for CEA. Mercaptoacetic acid–EDC–NHS reaction mechanism was used for anti-CEA immobilization. The effect of oxygen plasma treatment of the QCM electrode surface before bioreceptor preparation on the performance of the biosensor was tested and was found promising. CEA solutions with various concentrations were analysed using the bioreceptors to estimate the sensitivity and detection limit of the biosensor. The biosensors selectively recognized and captured CEA biomolecules with a detection limit of 0.06 and 0.09 ng mL−1 of CEA for oxygen plasma-treated (E2) and untreated (E1) bioreceptors, respectively. The sensitivity was estimated at 102 and 79 Hz, respectively, for E2 and E1. Clinical serum samples were analysed and the results were found in good agreement with the ELISA analysis. Long term stability was also found to be excellent. Langmuir adsorption isotherm was also conducted using the experimental results