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Multiplex bead binding assays using off-the-shelf components and common flow cytometers.

ABSTRACT: The ability to quantify protein-ligand interactions in an accurate and high-throughput manner is important in diverse areas of biology and medicine. Multiplex bead binding assays (MBBAs) are powerful methods that allow for simultaneous analysis of many protein-ligand interactions. Although there are a number of well-established MBBA platforms, there are few platforms suitable for research and development that offer rapid experimentation at low costs and without the need for specialized reagents or instruments dedicated for MBBA. Here, we describe a MBBA method that uses low-cost reagents and standard cytometers. The key innovation is the use of the essentially irreversible biotin-streptavidin interaction. We prepared a biotin-conjugated fluorescent dye and used it to produce streptavidin-coated magnetic beads that are labeled at distinct levels of fluorescence. We show the utility of our method in characterization of phage-displayed antibodies against multiple antigens of SARS-CoV-2, which substantially improves the throughput and dramatically reduces antigen consumption compared with conventional phage ELISA methods. This approach will make MBBAs more broadly accessible.

SUBMITTER: Hattori T

PROVIDER: S-EPMC7867647 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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Multiplex bead binding assays using off-the-shelf components and common flow cytometers.

Hattori Takamitsu T Koide Akiko A Panchenko Tatyana T Romero Larizbeth A LA Teng Kai Wen KW Corrado Alexis D AD Koide Shohei S

Journal of immunological methods 20201225

The ability to quantify protein-ligand interactions in an accurate and high-throughput manner is important in diverse areas of biology and medicine. Multiplex bead binding assays (MBBAs) are powerful methods that allow for simultaneous analysis of many protein-ligand interactions. Although there are a number of well-established MBBA platforms, there are few platforms suitable for research and development that offer rapid experimentation at low costs and without the need for specialized reagents ...[more]

PMID: 33358997

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Project description:Integrated serological surveillance (serosurveillance) involves testing for antibodies to multiple pathogens (or species) simultaneously and can be achieved using multiplex bead assays (MBAs). This systematic review aims to describe pathogens studied using MBAs, the operational implementation of MBAs, and how the data generated were synthesised. In November and December 2023, four databases were searched for studies utilising MBAs for the integrated serosurveillance of infectious diseases. Two reviewers independently screened and extracted data regarding the study settings and population, methodology, seroprevalence results, and operational implementation elements. Overall, 4765 studies were identified; 47 were eligible for inclusion, of which 41% (n = 19) investigated multiple malaria species, and 14% performed concurrent surveillance of malaria in combination with other infectious diseases (n = 14). Additionally, 14 studies (29%) investigated a combination of multiple infectious diseases (other than malaria), and seven studies examined a combination of vaccine-preventable diseases. Haiti (n = 8) was the most studied country, followed by Ethiopia (n = 6), Bangladesh (n = 3), Kenya (n = 3), and Tanzania (n = 3). Only seven studies were found where integrated serosurveillance was the primary objective. The synthesis of data varied and included the investigation of age-specific seroprevalence (n = 25), risk factor analysis (n = 15), and spatial analysis of disease prevalence (n = 8). This review demonstrated that the use of MBAs for integrated surveillance of multiple pathogens is gaining traction; however, more research and capabilities in lower- and middle-income countries are needed to optimise and standardise sample collection, survey implementation, and the analysis and interpretation of results. Geographical and population seroprevalence data can enable targeted public health interventions, highlighting the potential and importance of integrated serological surveillance as a public health tool.

Project description:BackgroundInfectious diseases continue to burden populations in Malaysia, especially among rural communities where resources are limited and access to health care is difficult. Current epidemiological trends of several neglected tropical diseases in these populations are at present absent due to the lack of habitual and efficient surveillance. To date, various studies have explored the utility of serological multiplex beads to monitor numerous diseases simultaneously. We therefore applied this platform to assess population level exposure to six infectious diseases in Sabah, Malaysia. Furthermore, we concurrently investigated demographic and spatial risk factors that may be associated with exposure for each disease.MethodsThis study was conducted in four districts of Northern Sabah in Malaysian Borneo, using an environmentally stratified, population-based cross-sectional serological survey targeted to determine risk factors for malaria. Samples were collected between September to December 2015, from 919 villages totaling 10,100 persons. IgG responses to twelve antigens of six diseases (lymphatic filariasis- Bm33, Bm14, BmR1, Wb123; strongyloides- NIE; toxoplasmosis-SAG2A; yaws- Rp17 and TmpA; trachoma- Pgp3, Ct694; and giardiasis- VSP3, VSP5) were measured using serological multiplex bead assays. Eight demographic risk factors and twelve environmental covariates were included in this study to better understand transmission in this community.ResultsSeroprevalence of LF antigens included Bm33 (10.9%), Bm14+ BmR1 (3.5%), and Wb123 (1.7%). Seroprevalence of Strongyloides antigen NIE was 16.8%, for Toxoplasma antigen SAG2A was 29.9%, and Giardia antigens GVSP3 + GVSP5 was 23.2%. Seroprevalence estimates for yaws Rp17 was 4.91%, for TmpA was 4.81%, and for combined seropositivity to both antigens was 1.2%. Seroprevalence estimates for trachoma Pgp3 + Ct694 were 4.5%. Age was a significant risk factors consistent among all antigens assessed, while other risk factors varied among the different antigens. Spatial heterogeneity of seroprevalence was observed more prominently in lymphatic filariasis and toxoplasmosis.ConclusionsMultiplex bead assays can be used to assess serological responses to numerous pathogens simultaneously to support infectious disease surveillance in rural communities, especially where prevalences estimates are lacking for neglected tropical diseases. Demographic and spatial data collected alongside serosurveys can prove useful in identifying risk factors associated with exposure and geographic distribution of transmission.

Dataset Information

Multiplex bead binding assays using off-the-shelf components and common flow cytometers.

Publications

Multiplex bead binding assays using off-the-shelf components and common flow cytometers.

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