Surface Plasmon Resonance Platform Research Articles

An Optical Device Based on a Chemical Chip and Surface Plasmon Platform for 2-Furaldehyde Detection in Insulating Oil.

2-Furaldehyde (2-FAL) is one of the main by-products of the degradation of hemicellulose, which is the solid material of the oil-paper insulating system of oil-filled transformers. For this reason, it has been suggested as a marker of the degradation of the insulating system; sensing devices for 2-FAL analysis in a wide concentration range are of high interest in these systems. An optical sensor system is proposed; this consists of a chemical chip, able to capture 2-FAL from the insulating oil, coupled with a surface plasmon resonance (SPR) probe, both realized on multimode plastic optical fibers (POFs). The SPR platform exploits gold nanofilm or, alternatively, a double layer of gold and silicon oxide to modulate the sensor sensitivity. The capturing chip is always based on the same molecularly imprinted polymer (MIP) as a receptor specific for 2-FAL. The system with the SPR probe based on a gold nanolayer had a higher sensitivity and a lower detection limit of fractions of μg L-1. Instead, the SPR probe, based on a double layer (gold and silicon oxide), has a lower sensitivity with a worse detection limit, and it is suitable for the detection of 2-FAL at concentrations of 0.01-1 mg L-1.

Highly sensitive detection of low-concentration sodium chloride solutions based on a gold-coated prism in Kretschmann setup

A gold-coated Kretschmann setup has been constructed and explored as a surface plasmon resonance (SPR) platform, specifically tailored for the detection of low-concentration sodium chloride (NaCl) solutions. The setup employs a BK7 prism coated with a 50 nm gold layer, serving as a plasmonic layer, to induce resonance. This resonance arises from the interplay between light waves and free electrons propagating at the interface of two media. The experimental findings reveal a notable resonance angle shift of 10° when the NaCl concentration is varied from 0 to 2.5 %. Furthermore, angle interrogation provides insightful details about the sensor's response to changes in the refractive index, showcasing a commendable sensitivity of 2400°/RIU, a high level of linearity at 0.9771, and an impressive resolution of 0.217 %. The demonstrated capabilities of this sensor underscore its potential for widespread applications, particularly in the monitoring of salt concentration across diverse domains such as seawater analysis, food processing, and fermentation processes. The robust performance and precision of this proposed sensor position it as a valuable tool with promising prospects for addressing the needs of various industries dependent on accurate salt concentration measurements.

Ammonia-trapping multilayer polymer urease film amplifier coated surface plasmon resonance sensor for ultra-sensitive urea detection

In this paper, we have developed a new urea signal amplification technology based on a multilayer urease polymer film self-assembly structure for the first time, enabling the realization of ultra-accurate urea optical detection. We demonstrate the effectiveness of this technique based on surface plasmon resonance (SPR) platform. When the urea signal amplifier is combined with SPR sensors, multilayer urease polymer film contacts with urea molecules, the urea molecules are oxidated into NH3 and CO2, causing the pH increase, which weakens the electrostatic interaction between different polymer layers and induces the RI reduction of the multilayer urease film, thereby causing the shift of the SPR spectrum. Experiment results confirm that urea sensitivity has been improved 12-fold, and the limit of detection (LOD) has been optimized to less than 10−3 mmol/L. Furthermore, the anti-interference has been improved 22.8-fold and is kept even the interference signal strength increases by 100 times. Moreover, multilayered urease polymer leads to the increase of urease on the sensor surface, thus expanding the detection range of urea by 3.5 times. This provides a new way to solve the contradiction between high sensitivity and large measurement range of optical fiber biosensor. Importantly, the concept of using enzyme-loaded polymer films for signal amplification can be adapted for the detection of various other biomolecules by replacing the enzyme with a specific recognition element for the target molecule. This opens doors for developing highly sensitive and specific biosensors for diverse applications in healthcare, environmental monitoring, and food safety.

3D-printed biosensors in biomedical applications exploiting plasmonic phenomena and antibody self-assembled monolayers.

In this work, a 3D-printed plasmonic chip based on a silver-gold bilayer was developed in order to enhance the optical response of the surface plasmon resonance (SPR) probe. More specifically, numerical and experimental results were obtained on the 3D-printed SPR platform based on a silver-gold bilayer. Then, the optimized probe's gold plasmonic interface was functionalized with a specific antibody directed against the p27Kip1 protein (p27), an important cell cycle regulator. The 3D-printed plasmonic biosensor was tested for p27 detection with good selectivity and a detection limit of 55 pM. The biosensor system demonstrated performance similar to commercially available ELISA (enzyme-linked immunoassay) kits, with several advantages, such as a wide detection range and a modular and simple-based architecture. The proposed biosensing technology offers flexible deployment options that are useful in disposable, low-cost, small-size, and simple-to-use biochips, envisaging future applications in experimental and biomedical research.

Immuno-SPR biosensor for the detection of Brucella abortus

A proof of principle biosensor for the Brucella abortus recognition onsite is presented. The system is based on a plasmonic optical fiber probe functionalized with an oriented antibody layer immobilized on a short polyethyleneglycol (PEG) interface through carbodiimide chemistry and protein G as an intermediate layer. The biosensor is inserted in a holder built in 3D printing technology, obtaining a custom holder useful for housing the sample to be measured and the equipment. The removable sensor chip is a low-cost Surface Plasmon Resonance (SPR) platform based on D-shaped plastic optical fibers (POFs), built-in in 3D printed connectors, used here for the first time to detect bacteria via a bio-receptor layer specific for its membrane protein. The performances of the biosensor in Brucella abortus recognition are tested by using two different SPR-POF probes combined with the same bio-receptor layer. The best sensor configuration has presented a sensitivity at low concentrations of one order of magnitude greater than the other. A limit of detection (LoD) of 2.8 bacteria/mL is achieved well competitive with other systems but without the need for amplification or special sample treatments. Specificity has been tested using Salmonella bacteria, and reproducibility, regenerability and stability are moreover evaluated. These experimental results pave the way for building an efficient and specific biosensor system for Brucella abortus detection onsite and in a few minutes. Moreover, the proposed POF-based SPR biosensor device, with respect to the already available technologies, could be a Point-of-care-test (POCT), simple to use, small-size and portable, low-cost, don’t necessary of a microfluidic system, and can be connected to the Internet (IoT).

Measuring ADAMTS-13 activity to diagnose thrombotic thrombocytopenic purpura: a novel, fast fiber-optic surface plasmon resonance immunoassay

BackgroundThrombotic thrombocytopenic purpura (TTP) is characterized by severe ADAMTS-13 activity deficiency (<10%). Diagnostic testing is challenging because of unavailability, high cost, and expert technician requirement of ADAMTS-13 enzyme assays. Cost-effective, automated fiber-optic surface plasmon resonance (FO-SPR) platforms show potential for developing diagnostic tests. Yet, FO-SPR has never been explored to measure enzymatic activities. ObjectivesTo develop an easy-to-use ADAMTS-13 activity assay utilizing optical fibers to rapidly diagnose TTP. MethodsThe ADAMTS-13 activity assay was designed and optimized using FO-SPR technology based on a previously described enzyme-linked immunosorbent assay setup. A calibration curve was generated to quantify ADAMTS-13 activity in plasma of healthy donors and patients with acute immune-mediated TTP (iTTP), hemolytic uremic syndrome, or sepsis. ADAMTS-13 activity data from FO-SPR and fluorescence resonance energy transfer-based strategies (FRETS)-VWF73 reference assays were compared. ResultsAfter initial assay development, optimization improved read-out magnitude and signal-to-noise ratio and reduced variation. Further characterization demonstrated a detection limit (6.8%) and inter-assay variation (Coefficient of variation, 7.2%) that showed good analytical sensitivity and repeatability. From diverse plasma samples, only plasma from patients with acute iTTP showed ADAMTS-13 activities below 10%. Strong Pearson correlation (r = 0.854) between FO-SPR and reference FRETS-VWF73 assays were observed for all measured samples. ConclusionsA fast ADAMTS-13 activity assay was designed onto automated FO-SPR technology. Optimization resulted in sensitive ADAMTS-13 activity measurements with a detection limit enabling clinical diagnosis of TTP within 3 hours. The FO-SPR assay proved strong correlation with the reference FRETS-VWF73 assay. For the first time, this assay demonstrated the capacity of FO-SPR technology to measure enzymatic activity in pre-clinical context.

A Novel Microchannel-in-a-Fiber Plasmonic Sensor

In this work, a novel Surface Plasmon Resonance (SPR) sensor has been designed, realized and investigated. The SPR platform was created by using a D-shaped plastic optical fiber shell (POFS), obtained by removing the graded index core region from the fiber’s protective jacket, in order to create a microchannel. The latter was then coated with a gold nanofilm in order to obtain the plasmonic sensitive region. The sensor’s structure includes an integrated measuring cell, being the solution under test injected into the microchannel using a peristaltic pump. The microchannel dimension (160 μm) allows to avoid microfluidic systems, typically adopted in this kind of SPR configurations. As a proof of concept, the plasmonic sensor has been tested as a refractometer using different water-ethanol solutions, with a refractive index range from 1.332 to 1.352. The experimental results have highlighted a sensitivity and a resolution equal to about 1026 nm/RIU and 1.95 × 10 <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">-4</sup> RIU, respectively. These values testify to the proposed sensor goodness and its suitability for biochemical sensing applications.

Estradiol Detection for Aquaculture Exploiting Plasmonic Spoon-Shaped Biosensors.

In this work, a surface plasmon resonance (SPR) biosensor based on a spoon-shaped waveguide combined with an estrogen receptor (ERα) was developed and characterized for the detection and the quantification of estradiol in real water samples. The fabrication process for realizing the SPR platform required a single step consisting of metal deposition on the surface of a polystyrene spoon-shaped waveguide featuring a built-in measuring cell. The biosensor was achieved by functionalizing the bowl sensitive surface with a specific estrogen receptor (ERα) that was able to bind the estradiol. In a first phase, the biosensor tests were performed in a phosphate buffer solution obtaining a limit of detection (LOD) equal to 0.1 pM. Then, in order to evaluate the biosensor's response in different real matrices related to aquaculture, its performances were examined in seawater and freshwater. The experimental results support the possibility of using the ERα-based biosensor for the screening of estradiol in both matrices.

Method for the Rapid Detection of SARS-CoV-2-Neutralizing Antibodies Using a Nanogel-Based Surface Plasmon Resonance Biosensor.

The efficacy of coronavirus disease 2019 (COVID-19) vaccination is closely related to the serum levels of SARS-CoV-2-neutralizing antibodies (NAb) that bind to the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. Therefore, the rapid and quantitative measurement of SARS-CoV-2 NAb in the sera of vaccinated individuals is essential to develop an effective vaccine and further achieve population immunity, that is, herd immunity. The plaque reduction neutralization test, the gold standard for NAb effectiveness in serological tests, is accurate but requires biosafety level 3 facilities because of the use of the virus, which hampers its application in common laboratories and clinical practice. Here, we developed a bioresponsive nanogel-based surface plasmon resonance (nSPR) platform that detects SARS-CoV-2 NAb in clinical samples without complicated pretreatment. We found that multivalent protein binding (MPB) between the nanogel-conjugated RBD protein and SARS-CoV-2 NAb yields significantly enhanced SPR signals compared to the nonspecific interference from serum proteins in the nSPR assay. The excellence of our nanogel-based SARS-CoV-2 NAb test is due to its selectivity for NAb, with resistance to all other proteins, allowing the rapid detection and quantification of NAbs in each individual. Importantly, this nSPR assay provides a NAb detection platform for easier and safer COVID-19 vaccination strategies.

Towards a point-of-care test to cover atto-femto and pico-nano molar concentration ranges in interleukin 6 detection exploiting PMMA-based plasmonic biosensor chips

Point-of-Care tests based on biomarkers, useful to monitor acute and chronic inflammations, are required for advances in medicine. In this scope, a key role is played by pro-inflammatory cytokines, of which interleukin 6 (IL-6) is generally thought as one of the most relevant. To use IL-6 in real scenarios, detection in ultra-low concentration ranges is required. In this work, two IL-6 biosensors are obtained by exploiting the combination of the same antibody self-assembled monolayer with two different plasmonic probes. This approach has demonstrated, via experimental results, that two different IL-6 concentration ranges can be explored. More specifically, IL-6 in an atto-femto molar range can be detected via polymer-based nanoplasmonic chips. On the other hand, a pico-nano molar range is obtained by a surface plasmon resonance platform in plastic optical fibers. As a proof of concept, the detection of IL-6 at the femto molar range has been obtained in Saliva and Serum. The results show that the proposed sensing approach could be useful in developing Point-of-Care devices based on a general setup with the capability to exploit both the plasmonic biosensor chips to monitor the IL-6 in the concentration range of interest, to provide an important support for the diagnosis and monitoring of oral and systemic diseases.

Detection of 2-Furaldehyde in Milk by MIP-Based POF Chips Combined with an SPR-POF Sensor.

An innovative optical-chemical sensor has been used to detect the 2-furaldehyde (2-FAL) in milk. The proposed sensing approach exploits the refractive index changing in a microstructured chip based on a plastic optical fiber (POF) with orthogonal micro-holes containing a specific molecularly imprinted polymer (MIP). This POF-MIP chemical chip modifies the surface plasmon resonance (SPR) phenomena excited in another sensor chip realized in POFs (SPR-POF) and connected in series. The proposed sensor configuration exploits MIP receptors avoiding any modification of the gold film of the SPR platform. This work reports the performance, particularly the high sensitivity and low detection limit, in complex matrices such as buffalo milk fortified with 2-FAL and in different commercial kinds of cow milk thermally treated for pasteurization. The measurements were carried out in about ten minutes by dropping the solution under-test on the planar D-shaped POF surface of the chemical chip. In contrast, on the gold surface of the SPR-POF platform, a water drop is always placed to excite the SPR phenomenon, which is modulated by the chemical chip via MIP-2-FAL binding. Furthermore, the experimental results demonstrated the pros and cons of the proposed sensor system. Thanks to the high sensitivity of the sensor system, the detection of 2-FAL in the diluted milk sample (1:50) was achieved. The dilution is required to reduce the interferent effect of the complex matrix.

A CRISPR/Cas12a-empowered surface plasmon resonance platform for rapid and specific diagnosis of the Omicron variant of SARS-CoV-2.

The outbreak of the COVID-19 pandemic was partially due to the challenge of identifying asymptomatic and presymptomatic carriers of the virus, and thus highlights a strong motivation for diagnostics with high sensitivity that can be rapidly deployed. On the other hand, several concerning SARS-CoV-2 variants, including Omicron, are required to be identified as soon as the samples are identified as ‘positive’. Unfortunately, a traditional PCR test does not allow their specific identification. Herein, for the first time, we have developed MOPCS (Methodologies of Photonic CRISPR Sensing), which combines an optical sensing technology-surface plasmon resonance (SPR) with the ‘gene scissors’ clustered regularly interspaced short palindromic repeat (CRISPR) technique to achieve both high sensitivity and specificity when it comes to measurement of viral variants. MOPCS is a low-cost, CRISPR/Cas12a-system-empowered SPR gene-detecting platform that can analyze viral RNA, without the need for amplification, within 38 min from sample input to results output, and achieve a limit of detection of 15 fM. MOPCS achieves a highly sensitive analysis of SARS-CoV-2, and mutations appear in variants B.1.617.2 (Delta), B.1.1.529 (Omicron) and BA.1 (a subtype of Omicron). This platform was also used to analyze some recently collected patient samples from a local outbreak in China, identified by the Centers for Disease Control and Prevention. This innovative CRISPR-empowered SPR platform will further contribute to the fast, sensitive and accurate detection of target nucleic acid sequences with single-base mutations.

A Plasmonic Biosensor Based on Light-Diffusing Fibers Functionalized with Molecularly Imprinted Nanoparticles for Ultralow Sensing of Proteins.

Plasmonic bio/chemical sensing based on optical fibers combined with molecularly imprinted nanoparticles (nanoMIPs), which are polymeric receptors prepared by a template-assisted synthesis, has been demonstrated as a powerful method to attain ultra-low detection limits, particularly when exploiting soft nanoMIPs, which are known to deform upon analyte binding. This work presents the development of a surface plasmon resonance (SPR) sensor in silica light-diffusing fibers (LDFs) functionalized with a specific nanoMIP receptor, entailed for the recognition of the protein human serum transferrin (HTR). Despite their great versatility, to date only SPR-LFDs functionalized with antibodies have been reported. Here, the innovative combination of an SPR-LFD platform and nanoMIPs led to the development of a sensor with an ultra-low limit of detection (LOD), equal to about 4 fM, and selective for its target analyte HTR. It is worth noting that the SPR-LDF-nanoMIP sensor was mounted within a specially designed 3D-printed holder yielding a measurement cell suitable for a rapid and reliable setup, and easy for the scaling up of the measurements. Moreover, the fabrication process to realize the SPR platform is minimal, requiring only a metal deposition step.

High-Throughput Surface Plasmon Resonance Biosensors for Identifying Diverse Therapeutic Monoclonal Antibodies.

Identification of antibodies targeting diverse functional epitopes on an antigen is highly crucial for discovering effective therapeutic candidates. Employing a traditional stepwise antibody "screening funnel" as well as prioritizing affinity-based selections over epitope-based selections, result in lead antibody panels lacking epitope diversity. In the present study, we employed an array-based surface plasmon resonance (SPR) platform to perform high-throughput epitope binning analysis on a large number of monoclonal antibodies (mAbs) generated in the early drug discovery process. The mAb panel contained clones from different antibody generation techniques and diverse transgenic mouse strains. The epitope binning results were analyzed in unique ways using various visualizations in the form of dendrograms and network plots, which assisted in determining diversity and redundancy in the mAb sample set. The binning data were further integrated with affinity information to evaluate the performance of seven different transgenic mouse strains. The combination of epitope binning results with binding kinetics and sequence analysis provided an effective and efficient way of selecting high affinity antibodies representing a diverse set of sequence families and epitopes.

In-situ and Real-Time Monitoring of the Interaction Between Lysins and Staphylococcus aureus Biofilm by Surface Plasmon Resonance.

Staphylococcus aureus can produce a multilayered biofilm embedded in extracellular polymeric matrix. This biofilm is difficult to remove, insensitive to antibiotics, easy to develop drug-resistant strains and causes enormous problems to environments and health. Phage lysin which commonly consists of a catalytic domain (CD) and a cell-wall binding domain (CBD) is a powerful weapon against bacterial biofilm. However, the real-time interaction between lysin and S. aureus biofilm is still not fully understood. In this study, we monitored the interactions of three lysins (ClyF, ClyC, PlySs2) against culture-on-chip S. aureus biofilm, in real-time, based on surface plasmon resonance (SPR). A typical SPR response curve showed that the lysins bound to the biofilm rapidly and the biofilm destruction started at a longer time. By using 1:1 binding model analysis, affinity constants (KD) for ClyF, ClyC, and PlySs2 were found to be 3.18 ± 0.127 μM, 1.12 ± 0.026 μM, and 15.5 ± 0.514 μM, respectively. The fact that ClyF and PlySs2 shared the same CBD but showed different affinity to S. aureus biofilm suggested that, not only CBD, but also CD affects the binding activity of the entire lysin. The SPR platform can be applied to improve our understanding on the complex interactions between lysins and bacterial biofilm including association (adsorption) and disassociation (destruction).

Enhanced Plasmonic Biosensor Utilizing Paired Antibody and Label-Free Fe3O4 Nanoparticles for Highly Sensitive and Selective Detection of Parkinson’s α-Synuclein in Serum

Parkinson’s disease (PD) is an acute and progressive neurodegenerative disorder, and diagnosis of the disease at its earliest stage is of paramount importance to improve the life expectancy of patients. -Synuclein (-syn) is a potential biomarker for the early diagnosis of PD, and there is a great need to develop a biosensing platform that precisely detects -syn in human body fluids. Herein, we developed a surface plasmon resonance (SPR) biosensor based on the label-free iron oxide nanoparticles (Fe3O4 NPs) and paired antibody for the highly sensitive and selective detection of -syn in serum samples. The sensitivity of the SPR platform is enhanced significantly by directly depositing Fe3O4 NPs on the Au surface at a high density to increase the decay length of the evanescent field on the Au film. Moreover, the utilization of rabbit-type monoclonal antibody (-syn-RmAb) immobilized on Au films allows the SPR platform to have a high affinity-selectivity binding performance compared to mouse-type monoclonal antibodies as a common bioreceptor for capturing -syn molecules. As a result, the current platform has a detection limit of /, which is 20,000-fold lower than that of commercial ELISA. The improved sensor chip can also be easily regenerated to repeat the -syn measurement with the same sensitivity. Furthermore, the SPR sensor was applied to the direct analysis of -syn in serum samples. By using a format of paired -syn-RmAb, the SPR sensor provides a recovery rate in the range from 94.5% to 104.3% to detect the -syn in diluted serum samples precisely. This work demonstrates a highly sensitive and selective quantification approach to detect -syn in human biofluids and paves the way for the future development in the early diagnosis of PD.

Gold nanoparticle enhanced multiplexed biosensing on a fiber optic surface plasmon resonance probe

We present an innovative multiplexing concept on a fiber optic surface plasmon resonance (FO-SPR) platform and demonstrate for the first time the simultaneous detection of two targets using the same FO sensor probe. Co(III)-NTA chemistry was used for oriented and stable co-immobilization of two different His6-tagged bioreceptors. T2C2 and MDTCS (i.e. fragments of the ADAMTS13 metalloprotease linked to the thrombotic thrombocytopenic purpura disorder) served as model system bioreceptors together with their respective targets (4B9 and II-1 antibodies). Gold nanoparticles were used here in an original way for discriminating the two targets in the same sample, in addition to their traditional signal amplification-role. After verifying the specificity of the selected model system, we studied the bioreceptor surface density and immobilization order. Innovative approach to lower the bioreceptor concentration below surface saturation resulted in an optimal detection of both targets, whereas the order of immobilization of the two bioreceptors did not give any significant difference. By sequentially immobilizing the T2C2 and MDTC bioreceptors, we established calibration curves in buffer and 100-fold diluted human blood plasma. This resulted in calculated limits of detection of 3.38 and 2.31 ng/mL in diluted plasma for 4B9 and II-1, respectively, indicating almost the same sensitivity as in buffer. Importantly, we also proved the applicability of the established calibration curves for quantifying the targets at random and more realistic ratios, directed by the design of experiments. This multiplexing study further expands the repertoire of applications on the FO-SPR biosensing platform, which together with its intrinsic features opens up great opportunities for diagnostics and life sciences.

SPR-Optical Fiber-Molecularly Imprinted Polymer Sensor for the Detection of Furfural in Wine.

A surface plasmon resonance (SPR) platform, based on a D-shaped plastic optical fiber (POF), combined with a biomimetic receptor, i.e., a molecularly imprinted polymer (MIP), is proposed to detect furfural (2-furaldheide, 2-FAL) in fermented beverages like wine. MIPs have been demonstrated to be a very convenient biomimetic receptor in the proposed sensing device, being easy and rapid to develop, suitable for on-site determinations at low concentrations, and cheap. Moreover, the MIP film thickness can be changed to modulate the sensing parameters. The possibility of performing single drop measurements is a further favorable aspect for practical applications. For example, the use of an SPR-MIP sensor for the analysis of 2-FAL in a real life matrix such as wine is proposed, obtaining a low detection limit of 0.004 mg L−1. The determination of 2-FAL in fermented beverages is becoming a crucial task, mainly for the effects of the furanic compounds on the flavor of food and their toxic and carcinogenic effect on human beings.

Calibration–free concentration analysis for quantification of anti-drug specific antibodies in polyclonal positive control antibodies and in clinical samples

Highly sensitive assays for anti–drug antibodies (ADAs) are both a regulatory requirement and requisite for proper evaluation of the effects of immunogenicity on clinical efficacy and safety. Determination of ADA assay sensitivity depends on positive control antibodies to represent naturally occurring or treatment–induced ADA responses. An accurate determination of the proportion of drug–specific antibodies in these polyclonal positive control batches is critical for correct evaluation of assay sensitivity.Target purification of positive control antibodies is commonly applied but infers the risk to lose a proportion of the antibodies. This may lead to an incorrect estimate of the ADA assay sensitivity, especially if high–affinity antibodies are lost that may be representative of natural ADAs with clinical implication.The Surface Plasmon Resonance platform on the Biacore™ systems offers methods for real–time analysis of biomolecular interactions without introducing any modifications to the analysed material. Calibration–free concentration analysis (CFCA) is such an application for determination of the proportion of drug–specific antibodies, which allows direct determination of active antibody concentrations, as defined by the ligand, in a flow–based system.Here, we present a novel CFCA method for ADA quantification developed and validated using polyclonal positive control antibodies against endogenous human insulin, insulin degludec (Tresiba®) and turoctocog alfa (NovoEight®).We find that CFCA precisely and accurately measures concentrations of drug–specific IgG antibodies with a precision of ±10% and 90%–112% recovery of expected values of monoclonal positive control antibodies. Additionally, we have achieved a more accurate measure of the sensitivity of a cell–based bioassay for in vitro neutralising ADAs using the specific concentration determined with CFCA. Moreover, we effectively quantified serum anti–insulin antibodies in high–titre clinical samples from individuals with diabetes mellitus. This application extends the relevance of the CFCA technology to analysis of immunogenicity for accurate quantification of ADAs in both the polyclonal positive control and in clinical samples.

An Automated Platform for Surface Plasmon Resonance-Based Sensors

This article presents the design of an automated platform to quickly, efficiently, and accurately manipulate the components required for a typical surface plasmon resonance (SPR) sensing instrument. In traditional SPR platforms, such parts are at prearranged and fixed positions. The proposed solution enables individual and group component movements by adjusting the alignment and positioning to ensure either the proper excitation and detection of the SPR. The platform's mechanical chassis is constructed with an aluminum V-slot profile and trapezoidal thread spindles and nuts to guide the components' displacement. An L-shaped frame specially designed for this purpose holds all the parts. Besides mechanical chassis, microcontrollers, motors, distance sensors, temperature sensors, and a touch-screen compose the automated platform components. The use of the so-called PPBIO biochip operating in the angular interrogation mode (AIM) demonstrates the platform's functioning. The obtained experimental results agree with the ones acquired with a similar but manually adjusted test platform.