Performance and validation of an adaptable multiplex assay for detection of serologic response to SARS-CoV-2 infection or vaccination

Kenny, Grace; Negi, Riya; O’Reilly, Sophie; Garcia-Leon, Alejandro; Alalwan, Dana; Gaillard, Colette Marie; Saini, Gurvin; Inzitari, Rosana; Feeney, Eoin; Yousif, Obada; Cotter, Aoife G.; Barra, Eoghan de; Sadlier, Corinna; Crispie, Fiona; Doran, Peter; Gautier, Virginie; Mallon, Patrick

doi:10.1016/j.jim.2022.113345

Cited by 13 publications

(4 citation statements)

References 22 publications

(16 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Few publications address the use of MSD multiplex technology for protein expression in the agriculture biotechnology industry . Nevertheless, there has been widespread use of this technology in the pharmaceutical industry and in clinical research. − MSD technology is very sensitive, with ultralow pico-gram level detection limits and a wide dynamic range of five or more orders of magnitude . This is particularly useful when analyzing samples with multiple traits that express trait proteins at widely varying levels.…”

Section: Description Of Techniquesmentioning

confidence: 99%

Immunoassays and Mass Spectrometry for Determination of Protein Concentrations in Genetically Modified Crops

Wang,

Bednarcik,

Ament

et al. 2024

J. Agric. Food Chem.

View full text Add to dashboard Cite

Quantifying protein levels in genetically modified (GM) crops is crucial in every phase of development, deregulation, and seed production. Immunoassays, particularly enzyme-linked immunosorbent assay, have been the primary protein quantitation techniques for decades within the industry due to their efficiency, adaptability, and credibility. Newer immunoassay technologies like Meso Scale Discovery and Luminex offer enhanced sensitivity and multiplexing capabilities. While mass spectrometry (MS) has been widely used for small molecules and protein detection in the pharmaceutical and agricultural industries (e.g., biomarkers, endogenous allergens), its use in quantifying protein levels in GM crops has been limited. However, as trait portfolios for GM crop have expanded, MS has been increasingly adopted due to its comparable sensitivity, increased specificity, and multiplexing capabilities. This review contrasts the benefits and limitations of immunoassays and MS technologies for protein measurement in GM crops, considering factors such as cost, convenience, and specific analytical needs. Ultimately, both techniques are suitable for assessing protein concentrations in GM crops, with MS offering complementary capabilities to immunoassays. This comparison aims to provide insights into selecting between these techniques based on the user's end point needs.

show abstract

Section: Description Of Techniquesmentioning

confidence: 99%

Immunoassays and Mass Spectrometry for Determination of Protein Concentrations in Genetically Modified Crops

Wang,

Bednarcik,

Ament

et al. 2024

J. Agric. Food Chem.

View full text Add to dashboard Cite

show abstract

“…Antibody response to nucleocapsid antigen during infection is shown to correlate with seropositivity 9 . Since the emergence of pandemic, 85 different serological test achieved authorization from FDA 11 . However, out of these 85 assays, majority of them are monoplex cut off assays and very few are fully quantitative assays.…”

Section: Introductionmentioning

confidence: 99%

Validation and Suitability Assessment of Multiplex Mesoscale Discovery Immunogenicity Assay for Establishing Serological Signatures Using Vaccinated, Non-vaccinated and Breakthrough SARS-COV-2 Infected Cases

Shengule,

Alai,

Bhandare

et al. 2024

Preprint

View full text Add to dashboard Cite

Antibody responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are multi-targeted and variable over time. Multiplex quantitative serological assays are needed to provide accurate and robust seropositivity data for establishment of serological signatures during vaccination and or infection. We describe here, validation and evaluation of an electro-chemiluminescence (ECL) based Mesoscale Discovery assays (MSD), for estimation of total and functional IgG to SARS-CoV-2 spike, nucleocapsid, and receptor binding (RBD) proteins in human serum samples to establish serological signatures of SARS-CoV-2 natural infection and breakthrough cases. The 9-PLEX assay was validated as per ICH, EMA and US FDA guidelines using a panel of sera samples including the NIBSC/WHO international reference panel (20/268). Assay demonstrated high specificity and selectivity in inhibition assays, wherein the homologous inhibition was more than 85% and heterologous inhibition was below 10 %. Assay also met predetermined acceptance criteria for precision (CV < 30 %), accuracy (70-130 %) and dilutional linearity. Method applicability to serological signatures was demonstrated using sera samples (N=45) representing vaccinated, infected and breakthrough cases. The method was able to establish distinct serological signatures and thus provide potential tool for seroprevalence of SARS-COV-2 during vaccination or infection.

show abstract

“…Additionally, it is now clear that not all Abs directed toward the Sars-2-CoV spike protein (C SP) are protective, necessitating the visualization of C SP sub-domain reactivity as well as the distribution of Abs across isotypes and toward new emerging variants. Although several platforms have been described to evaluate SARS-CoV-2-reactive Abs including [5][6][7][8][9][10][11][12], few of these methodologies are quantitative, making direct comparisons across platforms or the cohorts targeted in these studies difficult, thereby hampering our ability to define levels of Ab reactive with C SP or subdomains that correlate with protection. Such analyses are becoming increasingly important as SARS-CoV-2 variants resistant to prior vaccine responses continue to emerge [2].…”

Section: Introductionmentioning

confidence: 99%

A high-throughput multiplex array for antigen-specific serology with automated analysis

Rosenberg

Killian

Green

et al. 2023

Preprint

View full text Add to dashboard Cite

The utility of high-throughput systems to evaluate antigen-specific Ab has been highlighted by the SARS-CoV-2 pandemic. Pathogen specific antibody levels are often used to assess protection following vaccination and, in the case of novel pathogens, an indication of prior exposure. Several platforms exist to visualize antigen-specific Ab, however most are not quantitative and difficult to scale for population levels studies. Additionally, the sensitivity across platforms differs making direct comparisons between studies difficult. Cytometric Bead Arrays are an attractive platform for antigen-specific Ab measurements as they allow antibodies reactive against several antigens and of several isotypes to be performed simultaneously. Additionally, cytometric arrays exhibit a high sensitivity and can be designed to provide quantitative measurements. Using commercially available particles, a biotin-Streptavidin loading strategy, and the inclusion of indirect standards, we describe here a flexible system that can be modified to include a variety of antigens. We generated two arrays, one focused on b-Coronavirus antigens and one focused on Influenza. To support the high throughput capacity of this system, we developed a suit of automated tools to process raw data into antigen-reactive IgM, IgA, and IgG. We describe quality control requirements, assay performance, and normalizations to accurately quantitate antigen specific Ig.

show abstract

Performance and validation of an adaptable multiplex assay for detection of serologic response to SARS-CoV-2 infection or vaccination

Cited by 13 publications

References 22 publications

Immunoassays and Mass Spectrometry for Determination of Protein Concentrations in Genetically Modified Crops

Immunoassays and Mass Spectrometry for Determination of Protein Concentrations in Genetically Modified Crops

Validation and Suitability Assessment of Multiplex Mesoscale Discovery Immunogenicity Assay for Establishing Serological Signatures Using Vaccinated, Non-vaccinated and Breakthrough SARS-COV-2 Infected Cases

A high-throughput multiplex array for antigen-specific serology with automated analysis

Contact Info

Product

Resources

About