A novel immunoassay technique using principal component analysis for enhanced detection of emerging viral variants.

Rapid diagnostics are critical infectious disease tools that are designed to detect a known biomarker using antibodies specific to that biomarker. However, a way to detect unknown disease variants has not yet been achieved in a paper test format. We describe here a route to make an adaptable paper immunoassay that can detect an unknown biomarker, demonstrating it on SARS-CoV-2 variants.

Applications of Gold Nanoparticles in Plasmonic and Nanophotonic Biosensing.

The unique properties of plasmonic nanoparticles and nanostructures have enabled a broad range of applications in a diverse set of fields, ranging from biological sensing, cancer therapy, to catalysis. They have been some of the most studied nanomaterials due in part to their chemical stability and biocompatibility as well as supporting theoretical efforts. The synthesis and fabrication of plasmonic nanoparticles and nanostructures have now reached high precision and sophistication.

Gold Nanoparticle Paper Immunoassays for Sensing the Presence of in Oyster Hemolymph.

Seafood contamination with bacteria is a problem for aquaculture, especially with oysters, which are often consumed raw. Current methods for diagnosing bacterial pathogens in seafood involve lab-based assays such as polymerase chain reaction or culturing, which are time consuming and must occur in a centralized location. Detection of in a point-of-care assay would be a significant tool for food safety control measures.

The Influence of Preforming Protein Coronas on the Performance of Dengue NS1 Immunoassays.

The effect of preformed protein coronas on immunoassays for Dengue nonstructural protein 1 (NS1) immunoassays was investigated. The composition of the protein corona that forms around nanoparticle-antibody conjugates in human serum was characterized, and selected proteins from the corona were used for preformed coronas (human serum albumin and apolipoprotein A1). Coronas were formed and characterized by dynamic light scattering (DLS), and the nanoparticle-conjugate was probed by optical absorption spectroscopy.

Multiplexed rapid antigen tests developed using multicolored nanoparticles and cross-reactive antibody pairs: Implications for pandemic preparedness.

Global public health infrastructure is unprepared for emerging pathogen epidemics, in part because diagnostic tests are not developed in advance. The recent Zika, Ebola, and SARS-CoV-2 virus epidemics are cases in point. We demonstrate here that multicolored gold nanoparticles, when coupled to cross-reactive monoclonal antibody pairs generated from a single immunization regimen, can be used to create multiple diagnostics that specifically detect and distinguish related viruses.

Biophysical and biochemical insights in the design of immunoassays.

Background: Rapid antigen assays have been attractive for decentralized, point of care diagnostics because of their low cost, robustness, and ease of use. The development of a diagnostic assay for a newly emerging infectious disease needs to take into account the progression of a disease, whether there is human to human transmission, and patient biomarker levels with time, and these all impact the choice of antigen targets and affinity agents.

Scope Of Review: The factors involved in the biophysical design of rapid antigen immunoassays are discussed, focusing on antigen selection and designing for cross-reactivity.

Nanomaterial and interface advances in immunoassay biosensors.

Biosensors have been used for a remarkable array of applications, including infectious diseases, environmental monitoring, cancer diagnosis, food safety, and numerous others. In particular, the global COVID-19 pandemic has exposed a need for rapid tests, so the type of biosensor that has gained considerable interest recently are immunoassays, which are used for rapid diagnostics. The performance of paper-based lateral flow and dipstick immunoassays is influenced by the physical properties of the nanoparticles (NPs), NP-antibody conjugates, and paper substrate.

Correction: SARS-CoV-2 and approaches for a testing and diagnostic strategy.

Correction for 'SARS-CoV-2 and approaches for a testing and diagnostic strategy' by Delyan R. Hristov , , 2021, , 8157-8173, DOI: https://doi.org/10.

SARS-CoV-2 and approaches for a testing and diagnostic strategy.

The COVID-19 pandemic has led to an unprecedented global health challenge, creating sudden, massive demands for diagnostic testing, treatment, therapies, and vaccines. In particular, the development of diagnostic assays for SARS-CoV-2 has been pursued as they are needed for quarantine, disease surveillance, and patient treatment. One of the major lessons the pandemic highlighted was the need for fast, cheap, scalable and reliable diagnostic methods, such as paper-based assays.

Developing a Paper-Based Antigen Assay to Differentiate between Coronaviruses and SARS-CoV-2 Spike Variants.

COVID-19 first appeared in December of 2019 in Wuhan, China. Since then, it has become a global pandemic. A robust and scalable diagnostics strategy is crucial for containing and monitoring the pandemic.

Optimization of paper-based nanoparticle immunoassays for direct detection of the bacterial pathogen in oyster hemolymph.

The detection of foodborne pathogens is critical for disease control and infection prevention, especially in seafood consumed raw or undercooked. Paper-based diagnostic tools are promising for rapid fieldable detection and provide a readout by eye due to the use of gold nanoparticle immunoprobes. Here we study different strategies to overcome these challenges in a real biological matrix, oyster hemolymph, for the detection of the pathogenic bacteria Vibrio parahaemolyticus (Vp).

The Immunoprobe Aggregation State is Central to Dipstick Immunoassay Performance.

As new infectious disease outbreaks become more likely, it is important to be able to develop and deploy appropriate testing in time. Paper-based immunoassays are rapid, cheap, and easy to produce at scale and relatively user friendly but often suffer from low selectivity and sensitivity. Understanding the molecular mechanisms of paper immunoassays may help improve and hasten development and therefore production and market availability.

Repurposing Old Antibodies for New Diseases by Exploiting Cross-Reactivity and Multicolored Nanoparticles.

We exploit the cross-reactivity of dengue (DENV) and Zika (ZIKV) virus polyclonal antibodies for nonstructural protein 1 (NS1) to construct a selective sensor that can detect yellow fever virus (YFV) NS1 in a manner similar to chemical olfaction. DENV and ZIKV antibodies were screened for their ability to bind to DENV, ZIKV, and YFV NS1 by enzyme linked immunosorbent assay (ELISA) and in pairs in paper immunoassays. A strategic arrangement of antibodies immobilized on paper and conjugated to different colored gold NPs was used to distinguish the three biomarkers.

Distributed Biological Foundries for Global Health.

Historically, many industries such as manufacturing have undergone a trend away from centralized, large-scale production toward a more distributed form. Currently, this same trend is witnessed in biological manufacturing and bioprocessing, with the rise of biological foundries where one can synthesize, grow, isolate, and purify a broad range of biologics. The adoption of distributed practices for biological processing has significant implications for healthcare, diagnostics, and therapies.

Detection of resistance protein A (MxA) in paper-based immunoassays with surface enhanced Raman spectroscopy with AuAg nanoshells.

Myxovirus protein A (MxA) is a biomarker that can be used to distinguish between viral and bacterial infections. While MxA lateral flow assays (LFAs) have been successfully used for viral vs. bacterial differential diagnosis for children, the clinically relevant level of MxA for adults has been reported to be 100 times lower, which is too low for traditional LFAs.

Protease Degradation of Protein Coronas and Its Impact on Cancer Cells and Drug Payload Release.

The effect of matrix metalloproteinases (MMPs) on preformed protein coronas around spherical gold nanoparticles (AuNPs) was studied. Protein coronas of different compositions (human serum, human serum albumin, and collagen IV) were formed around AuNPs and characterized. The protease MMP-9 had different effects on the corona depending on the corona composition, resulting in different changes to the corona hydrodynamic diameter ( D).

PERSIA for Direct Fluorescence Measurements of Transcription, Translation, and Enzyme Activity in Cell-Free Systems.

Quantification of biology's central dogma (transcription and translation) is pursued by a variety of methods. Direct, immediate, and ongoing quantification of these events is difficult to achieve. Common practice is to use fluorescent or luminescent proteins to report indirectly on prior cellular events, such as turning on a gene in a genetic circuit.

Designing Paper-Based Immunoassays for Biomedical Applications.

Paper-based sensors and assays have been highly attractive for numerous biological applications, including rapid diagnostics and assays for disease detection, food safety, and clinical care. In particular, the paper immunoassay has helped drive many applications in global health due to its low cost and simplicity of operation. This review is aimed at examining the fundamentals of the technology, as well as different implementations of paper-based assays and discuss novel strategies for improving their sensitivity, performance, or enabling new capabilities.

Reporter Selection for Nanotags in Multiplexed Surface Enhanced Raman Spectroscopy Assays.

We report a quantitative evaluation of the choice of reporters for multiplexed surface-enhanced Raman spectroscopy (SERS). An initial library consisted of 15 reporter molecules that included commonly used Raman dyes, thiolated reporters, and other small molecules. We used a correlation matrix to downselect Raman reporters from the library to choose five candidates: 1,2-bis(4-pyridyl)ethylene, 4-mercaptobenzoic acid, 3,5-dichlorobenzenthiol, pentachlorothiophenol, and 5,5'-dithiobis(2-nitrobenzoic acid).

Ampli: A Construction Set for Paperfluidic Systems.

The design and fabrication of reconfigurable, modular paperfluidics driven by a prefabricated reusable block library, asynchronous modular paperfluidic linear instrument-free (Ampli) block, are reported. The blocks are inspired by the plug-and-play modularity of electronic breadboards that lower prototyping barriers in circuit design. The resulting biochemical breadboard is a paperfluidic construction set that can be functionalized with chemical, biological, and electrical elements.

Physical Properties of Biomolecules at the Nanomaterial Interface.

The unique size and material dependent properties of nanoparticles have made them highly attractive for biological and medical applications. However, combining nanoparticles with biomolecules and biological environments has faced many challenges. These interface issues often involve protein denaturation, steric hindrance, and orientational issues for the biomolecule, which can impair function and decrease overall performance of the nanoparticle-biomolecule conjugate.

Challenges of the Nano-Bio Interface in Lateral Flow and Dipstick Immunoassays.

Lateral flow assays (LFAs) are highly attractive for point-of-care (POC) diagnostics for infectious disease, food safety, and many other medical uses. The unique optical, electronic, and chemical properties that arise from the nanostructured and material characteristics of nanoparticles provide an opportunity to increase LFA sensitivity and impart novel capabilities. However, interfacing to nanomaterials in complex biological environments is challenging and can result in undesirable side effects such as non-specific adsorption, protein denaturation, and steric hindrance.

Design of SERS nanotags for multiplexed lateral flow immunoassays.

Surface enhanced Raman spectroscopy (SERS) has been attractive for enhancing the sensitivity of lateral flow immunoassays (LFA). A format that has enabled specific detection of biomarkers is to use Raman reporter molecules linked to gold nanoparticles (NPs), which are conjugated to antibodies specific for the target of interest. Many factors such as the NP and Ab properties and the method of signal readout impact the sensitivity of a SERS based immunoassay.