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Magplex beads

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MagPlex beads are magnetic microspheres used in various laboratory applications. They serve as a solid support for the immobilization and manipulation of biomolecules, facilitating separation and purification processes.

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Lab products found in correlation

Flexmap 3d instrument, by DiaSorin (7 mentions) Magpix, by DiaSorin (5 mentions) Goat anti human igg pe, by Southern Biotech (4 mentions) Sulfo n hydroxysulfosuccinimide, by Thermo Fisher Scientific (3 mentions) 1 ethyl 3 3 dimethylaminopropyl carbodiimide, by Thermo Fisher Scientific (3 mentions) Ebov soluble gp, by IBT Bioservices (2 mentions) Recombinant ebov gp, by IBT Bioservices (2 mentions) Flexmap 3d, by DiaSorin (2 mentions) Goat anti human iga pe, by Southern Biotech (2 mentions) Prism 9, by GraphPad (2 mentions) Luminex magpix system, by DiaSorin (2 mentions) Bovine serum albumin (bsa), by Genecust (2 mentions) Bovine serum albumin (bsa), by DiaSorin (2 mentions) Xmap antibody coupling kit, by DiaSorin (2 mentions) Ebov gp, by IBT Bioservices (1 mentions) 200 analyser, by DiaSorin (1 mentions) Xmap sars cov 2 multi antigen igg assay, by DiaSorin (1 mentions) Sheath fluid, by DiaSorin (1 mentions) R phycoerythrin labeled goat anti human igg, by Jackson ImmunoResearch (1 mentions) 96 well plate, by Corning (1 mentions)

38 protocols using magplex beads

1

Covalent Coupling of SARS-CoV-2 S Protein to Beads

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Proteins were covalently coupled to Magplex beads (Luminex Corporation) using a two-step carbodiimide reaction and a ratio of 75 μg protein SARS-CoV-2 S protein to 12,5 million beads. Magplex beads (Luminex Corporation) were washed with 100 mM monobasic sodium phosphate pH 6.2, activated for 30 min on a rotor at RT by addition of Sulfo-N-Hydroxysulfosuccinimide (Thermo Fisher Scientific) and 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (Thermo Fisher Scientific). The activated beads were washed three times with 50 mM MES pH 5.0 and added to SARS-CoV-2 S protein which was diluted in 50 mM MES pH 5.0. The beads and protein were incubated for 3 h on a rotator at RT. Afterward, the beads were washed with PBS and blocked with PBS containing 0.1% BSA, 0.02% Tween-20 and 0.05% Sodium Azide at pH 7.0 for 30 min on a rotator at RT. Finally, the beads were washed and stored in PBS containing 0.05% Sodium Azide at 4°C and used within 3 months.
Brouwer P.J., Brinkkemper M., Maisonnasse P., Dereuddre-Bosquet N., Grobben M., Claireaux M., de Gast M., Marlin R., Chesnais V., Diry S., Allen J.D., Watanabe Y., Giezen J.M., Kerster G., Turner H.L., van der Straten K., van der Linden C.A., Aldon Y., Naninck T., Bontjer I., Burger J.A., Poniman M., Mykytyn A.Z., Okba N.M., Schermer E.E., van Breemen M.J., Ravichandran R., Caniels T.G., van Schooten J., Kahlaoui N., Contreras V., Lemaître J., Chapon C., Fang R.H., Villaudy J., Sliepen K., van der Velden Y.U., Haagmans B.L., de Bree G.J., Ginoux E., Ward A.B., Crispin M., King N.P., van der Werf S., van Gils M.J., Le Grand R, & Sanders R.W. (2021). Two-component spike nanoparticle vaccine protects macaques from SARS-CoV-2 infection. Cell, 184(5), 1188-1200.e19.
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2

Covalent Coupling of SARS-CoV-2 S Protein to Beads

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Proteins were covalently coupled to Magplex beads (Luminex Corporation) via a two-step carbodiimide reaction using a ratio of 75 μg SARS-CoV-2 S to 12,5 million beads. Magplex beads (Luminex Corporation) were washed with 100 mM mono-basic sodium phosphate pH 6.2 and activated for 30 min on a rotor at RT by addition of Sulfo-N-Hydroxysulfosuccinimide (Thermo Fisher Scientific) and 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (Thermo Fisher Scientific). The activated beads were washed three times with 50 mM MES pH 5.0 and added to SARS-CoV-2 S protein, which was diluted in 50 mM MES pH 5.0. The coupling reaction was incubated for 3 h on a rotator at RT. The beads were subsequently washed with PBS and blocked with PBS containing 2% BSA, 3% FCS and 0.02% Tween-20 for 30 min on a rotator at RT. Finally, the beads were washed and stored in PBS containing 0.05% Sodium Azide at 4°C and used within 3 months.
Sulbaran G., Maisonnasse P., Amen A., Effantin G., Guilligay D., Dereuddre-Bosquet N., Burger J.A., Poniman M., Grobben M., Buisson M., Dergan Dylon S., Naninck T., Lemaître J., Gros W., Gallouët A.S., Marlin R., Bouillier C., Contreras V., Relouzat F., Fenel D., Thepaut M., Bally I., Thielens N., Fieschi F., Schoehn G., van der Werf S., van Gils M.J., Sanders R.W., Poignard P., Le Grand R, & Weissenhorn W. (2022). Immunization with synthetic SARS-CoV-2 S glycoprotein virus-like particles protects macaques from infection. Cell Reports Medicine, 3(2), 100528.
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3

EBOV GP and sGP Multiplex Assay

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Recombinant EBOV GP and EBOV soluble GP (sGP; IBT Bioservices) were coupled to MagPlex beads (Luminex) via sulfo-NHS coupling chemistry. Heat-inactivated samples were diluted 1:50 in 1X PBS + 0.1% bovine serum albumin (BSA) + 0.05% Tween20 and incubated with antigen-coupled beads for 2h. Beads were washed, and secondary antibodies specific for non-human primate antibody subclasses IgG1 (clone 7H11; NHP Resource Center), IgG2 (clone 3C10; NHP Resource Center), IgG3 (clone 2G11; NHP Resource Center) and isotypes (IgM, IgA) were detected by incubating with 0.65μg/ml of secondary antibodies (Southern Biotech) for 1h at room temperature. Beads were washed 6X with assay buffer and incubated with a PE-labeled anti-mouse IgG1 (1μg/ml) for 1 hour at room temperature. Beads were washed analyzed on a Flexmap 3D instrument (Luminex). The median fluorescent intensity of 30 beads/region was recorded. Sera from naive cynomolgus macaques were used to establish baseline reactivity and thresholds for positivity.
Gunn B.M., McNamara R.P., Wood L., Taylor S., Devadhasan A., Guo W., Das J., Nilsson A., Shurtleff A., Dubey S., Eichberg M., Suscovich T.J., Saphire E.O., Lauffenburger D., Coller B.A., Simon J.K, & Alter G. (2023). Antibodies against the Ebola virus soluble glycoprotein are associated with long-term vaccine-mediated protection of non-human primates. Cell reports, 42(4), 112402.
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4

Rhesus Macaque FcγR Binding Assay

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Recombinant EBOV GP or EBOV soluble GP (sGP; IBT Bioservices) were coupled to MagPlex beads (Luminex) via sulfo-NHS coupling chemistry. Three activating FcγR receptors (FcγR2A, FcγR3A, FcγR3B) and one inhibitory receptor (FcγR2B). NHP have polymorphic receptors and among the four rhesus FcγR2A receptors, only FcγR2A-2 and FcγR2A-3 show similar IgG binding, and within the three known FcγR3A receptors (-1, -2, -3) (69 (link)), FcγR3A-1 and FcγR3A-3 have distinct IgG binding profiles, and thus binding antibodies to rhesus FcγR2A-3, FcγR3A-1 and FcγR3A-3 was measured. Heat-inactivated samples were diluted 1:50 (rhesus FcRs) or 1:500 (human FcRs) in 1X PBS + 0.1% bovine serum albumin (BSA) + 0.05% Tween20 and incubated with antigen-coupled beads for 2h. Beads were washed and incubated with recombinant biotinylated Fc-receptors that were tetramerized via Streptavidin-PE for 1 hour at room temperature. Beads were washed analyzed on a Flexmap 3D instrument (Luminex). The median fluorescent intensity of 30 beads/region was recorded. Sera from naive cynomolgus macaques were used to establish baseline reactivity and thresholds for positivity.
Gunn B.M., McNamara R.P., Wood L., Taylor S., Devadhasan A., Guo W., Das J., Nilsson A., Shurtleff A., Dubey S., Eichberg M., Suscovich T.J., Saphire E.O., Lauffenburger D., Coller B.A., Simon J.K, & Alter G. (2023). Antibodies against the Ebola virus soluble glycoprotein are associated with long-term vaccine-mediated protection of non-human primates. Cell reports, 42(4), 112402.
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5

Luminex Assay for COVID-19 Antibody Detection

Cited 1 time
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An explorative Luminex assay was developed to investigate antigen-specific IgG and IgA in serum and saliva. A recombinant prefusion ectodomain trimer of SARS-CoV-2 S protein, the monomeric RBD of the S protein, and N protein were designed, produced, and purified as previously described (42 (link), 43 (link)). The proteins were covalently coupled to Magplex beads (Luminex) using a two-step carbodiimide reaction at a ratio of 75 μg protein to 12.5 million beads for S, at an equimolar concentration for N, and at 3× the equimolar concentration for RBD. Beads were washed with 100 mM monobasic sodium phosphate, pH 6.2, activated with sulfo-N-hydroxysulfosuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (Thermo Fisher Scientific), and incubated for 30 min on a rotator at room temperature (RT). Activated beads were washed 3 times with 50 mM MES (morpholineethanesulfonic acid), pH 5.0, proteins were added, and beads were incubated for 3 h on a rotator at RT. The beads were washed with phosphate-buffered saline (PBS) and blocked with PBS containing 2% bovine serum albumin (BSA), 3% fetal calf serum, and 0.02% Tween 20 at pH 7.0 (PBS-blocking) for 30 min on a rotator at RT. Finally, the beads were washed, stored in PBS containing 0.05% sodium azide at 4°C, and used within 6 weeks.
Keuning M.W., Grobben M., de Groen A.E., Berman-de Jong E.P., Bijlsma M.W., Cohen S., Felderhof M., de Groof F., Molanus D., Oeij N., Rijpert M., van Eijk H.W., Koen G., van der Straten K., Oomen M., Visser R., Linty F., Steenhuis M., Vidarsson G., Rispens T., Plötz F.B., van Gils M.J, & Pajkrt D. (2021). Saliva SARS-CoV-2 Antibody Prevalence in Children. Microbiology Spectrum, 9(2), e00731-21.
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6

Multiplex Luminex SARS-CoV-2 IgG Assay

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The xMAP SARS-CoV-2 Multi-Antigen IgG Assay (Luminex, Texas; United States) Research Use Only (RUO) kit version 1.00 was used. This assay tests for IgG antibodies in human serum against the SARS-CoV-2 antigens: N, RBD and S1 (Supplementary Material A table 3). The assay is a Luminex multiplex assay using Luminex MagPlex beads. The assay was completed following the manufacturer's instructions for use package insert (Supplementary Material A.3). The 50 samples were tested in a 96 well round-bottomed plate and positive and negative control samples provided in the test kit were included. The plate was then read using the Luminex 200 analyser with a provided xPONENT software protocol.
Individual sample results were considered acceptable when: IgG control bead determined as ‘present’ (MFI > 2500 call threshold), IgA and IgM control beads determined as ‘absent’ (MFI < call threshold determined by software) and background bead determined as ‘passed’ (MFI < 700 call threshold). Samples were considered SARS-CoV-2 IgG antibody positive when their MFI values for both Nucleocapsid bead and RBD bead were > 700 call threshold for each bead. Each run was considered acceptable when both negative and positive control samples met individual sample requirements as above and provided negative and positive SARS-CoV-2 IgG results respectively.
Cox A., Stevens M., Kallon D., Gupta A, & White E. (2023). Comparative evaluation of Luminex based assays for detection of SARS-CoV-2 antibodies in a transplantation laboratory. Journal of Immunological Methods, 517, 113472.
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7

Antigen Coupling for Multiplex Serology

Cited 2 times
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We coupled antigens to spectrally distinct MagPlex beads (Luminex, https://www.luminexcorp.com) by using EDC/s-NHS coupling for all standard (MULTICOV-AB) antigens (14 (link)). We coupled receptor-binding domains (RBDs) from VOCs by using Anteo coupling (AnteoTech, https://www.anteotech.com) according to the manufacturer’s instructions (15 (link)).
Dulovic A., Strengert M., Ramos G.M., Becker M., Griesbaum J., Junker D., Lürken K., Beigel A., Wrenger E., Lonnemann G., Cossmann A., Stankov M.V., Dopfer-Jablonka A., Kaiser P.D., Traenkle B., Rothbauer U., Krause G., Schneiderhan-Marra N, & Behrens G.M. (2022). Diminishing Immune Responses against Variants of Concern in Dialysis Patients 4 Months after SARS-CoV-2 mRNA Vaccination. Emerging Infectious Diseases, 28(4), 743-750.
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8

Coupling of SARS-CoV-2 Antigens to Beads

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Coupling of antigens to spectrally distinct MagPlex beads (Cat #MC10XXX-01, Luminex Corporation, USA) was done by EDC/s-NHS coupling for all standard MULTICOV-AB antigens [26] (link). Receptor-binding domains (RBD)s from VoC were coupled using Anteo coupling (Cat #A-LMPAKMM-10, Anteo Tech Reagents, Australia) following the manufacturer's instructions [27] (link).
Strengert M., Becker M., Ramos G.M., Dulovic A., Gruber J., Juengling J., Lürken K., Beigel A., Wrenger E., Lonnemann G., Cossmann A., Stankov M.V., Dopfer-Jablonka A., Kaiser P.D., Traenkle B., Rothbauer U., Krause G., Schneiderhan-Marra N, & Behrens G.M. (2021). Cellular and humoral immunogenicity of a SARS-CoV-2 mRNA vaccine in patients on haemodialysis. EBioMedicine, 70, 103524.
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9

Quantitative ZIKV and DENV-VLP Binding Assay

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The Luminex assay was conducted by FlexMap 3D (Luminex, Austin, TX, USA), and the conjugation of VLP was previously reported (70 (link)). Briefly, 65 μg ZIKV and DENV-VLP was conjugated to 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride, ECD/N-hydroxy-sulfo-succinimide, NHS (Thermo Fisher, Waltham, MA, USA), and activated in 12.5 million MagPlex beads (Luminex, Austin, TX, USA) in 50 mM 2-(N-morpholino)ethanesulfonic acid buffer, pH 7.0 or 6.0, for 120 min at room temperature. After conjugation, excess active residues were blocked by sample buffer (1% bovine serum albumin [BSA] in d-PBS) overnight at 4°C. A total of 10,000 ZIKV- and DENV-VLP conjugated beads/mL and anti-ZIKV MAb was incubated at room temperature in sample buffer for 90 min and washed with phosphate-buffered saline plus 0.05% Tween 20 (PBST). After washing, the beads were incubated with 10 μg/mL phycoerythrin-labeled anti-rabbit IgG (Thermo Fisher, Waltham, MA, USA) for 60 min. The beads were washed and mixed with sheath fluid (Luminex, Austin, TX, USA). The plates were measured the fluorescence intensity by FlexMap 3D.
Tsuji I., Vang F., Dominguez D., Karwal L., Sanjali A., Livengood J.A., Davidson E., Fouch M.E., Doranz B.J., Das S.C, & Dean H.J. (2022). Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function. Journal of Virology, 96(11), e00071-22.
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10

Luminex-Based Antibody Detection

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Antibody levels in serum and milk were assessed using a custom Luminex assay similar to previously described (29 ). In short, viral antigens were covalently coupled to Luminex Magplex beads with a two-step carbodiimide reaction at a ratio of 37 μg protein to 12.5 million beads for RSV-F glycoprotein and equimolar amounts for other proteins. Following the outcome of optimization experiments, serum was diluted 1:10,000, and milk was diluted 1:100. Beads and diluted samples were incubated overnight, followed by detection with goat-anti-human IgG-PE or goat-anti-human IgA-PE (Southern Biotech). Read-out was performed on a Magpix (Luminex). The resulting median fluorescence intensity (MFI) values are the median of at least 50 beads per well and were corrected by subtraction of MFI values from the buffer and beads-only wells.
Grobben M., Juncker H.G., van der Straten K., Lavell A.H., Schinkel M., Buis D.T., Wilbrink M.F., Tejjani K., Claireaux M.A., Aartse A., de Groot C.J., Pajkrt D., Bomers M.K., Sikkens J.J., van Gils M.J., van Goudoever J.B, & van Keulen B.J. (2022). Decreased Passive Immunity to Respiratory Viruses through Human Milk during the COVID-19 Pandemic. Microbiology Spectrum, 10(4), e00405-22.
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