Luminex 200 platform

Manufactured by Bio-Rad

Sourced in United States, United Kingdom

The Luminex 200 platform is a multiplex assay system that utilizes flow cytometry technology to measure multiple analytes simultaneously in a single sample. The system employs color-coded magnetic beads, each coated with a specific capture antibody, to enable the detection and quantification of various proteins, cytokines, or other biomolecules in a sample.

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Lab products found in correlation

Bio plex manager software, by Bio-Rad (2 mentions) Biorad bio plex software, by Bio-Rad (2 mentions) Fluorokine multianalyte profiling kit, by R&D Systems (2 mentions) Multiplex luminometry, by Bio-Rad (1 mentions) Sandwich elisa, by Thermo Fisher Scientific (1 mentions) Solid phase sandwich elisa, by R&D Systems (1 mentions) Multiplex kits, by R&D Systems (1 mentions) Milliplex map human cytokine chemokine kit, by Merck Group (1 mentions) Milliplex multiplex assay, by Merck Group (1 mentions) Bio plex manager 6, by Bio-Rad (1 mentions) Procartaplex multiplex immunoassay, by Thermo Fisher Scientific (1 mentions) Bio plex manager v6, by Bio-Rad (1 mentions) Bio plex manager software version 6, by Bio-Rad (1 mentions) Bio plex pro human cytokine screening panel, by Bio-Rad (1 mentions)

23 protocols using luminex 200 platform

Multiplex Immune Profiling of Uveal and Cutaneous Melanoma

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Sera from uveal and CM patients were assayed to evaluate the levels of cytokines and soluble immune checkpoint molecules (sICs) by multiplex immunoassay analysis using the ProcartaPlex Human Inflammation Panel (20 Plex, catalog number EPX200-12185-901; sE-Selectin; GM-CSF; ICAM-1/CD54; IFN alpha; IFN gamma; IL-1 alpha; IL-1 beta; IL-4; IL-6; IL-8; IL-10; IL-12p70; IL-13; IL-17A/CTLA-8; IP-10/CXCL10; MCP-1/CCL2; MIP-1alpha/CCL3; MIP-1 beta/CCL4; sP-Selectin; TNF alpha) (eBioscence) and the Human Immuno-Oncology Checkpoint 14-plex ProcartaPlex Panel 1 (catalog number EPX14A-15803-901; BTLA; GITR; HVEM; IDO; LAG-3: 47; PD-1; PD-L1; PD-L2; TIM-3; CD28; CD80; CD137; CD27; CD152) (eBioscence). Assay was conducted using 50 µl of serum for each sample and adding it in a 96-well plate with a mixture of color-coded magnetic beads coated with antibody that recognize specific analytes. Later, biotinylated detection antibodies that bind analytes of interest were added and then bound to Phycoerythrin-conjugated streptavidin that through its signal intensity allow to detect the analyte concentration. Samples were measured using Luminex 200 platform (BioPlex, Bio-Rad) and data, expressed in pg/ml of protein, were analyzed using Bio-Plex Manager Software.

Rossi E., Zizzari I.G., Di Filippo A., Acampora A., Pagliara M.M., Sammarco M.G., Simmaco M., Lionetto L., Botticelli A., Bria E., Marchetti P., Blasi M.A., Tortora G., Schinzari G, & Nuti M. (2022). Circulating immune profile can predict survival of metastatic uveal melanoma patients: results of an exploratory study. Human Vaccines & Immunotherapeutics, 18(3), 2034377.

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Cord Blood Biomarker Profiling for EOS

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Nine APR proteins were measured in cord blood: PCT, CRP, haptoglobin (Hp), ferritin, fibrinogen, tissue plasminogen activator, serum amyloid A (SAA), serum amyloid P (SAP), and α-2-macroglobulin. These markers were selected upon careful review of the literature for potentially promising markers of EOS and available platforms from commercially available kits [7 (link), 32 (link)–39 (link)]. The proteins were measured in cord blood plasma in duplicate using magnetic bead-based immunoassays (Bio-Plex Pro^TM Human Acute Phase Multiplex Assays [4-plex (α-2-macroglobulin, CRP, Hp, SAP) and 5-plex (ferritin, fibrinogen, PCT, SAA, tissue plasminogen activator)]; Bio-Rad Laboratories Inc.; Hercules, CA). Samples were thawed on ice and prepared in 1:1000 and 1:100 dilution for the respective panels. The kits were run on a Luminex 200 platform with Bio-Plex Manager 6.1 and Milliplex Analyst 5.1 software used for analysis. A 5 parameter logistic regression was used for standard curve fitting. Coefficient of variation for biomarkers were 0.53%-1.81%. The samples were prepared and assays were performed in the Comprehensive Metabolic Core at Northwestern University. Turnaround time to obtain results from this assay is 8 hours.

Mithal L.B., Palac H.L., Yogev R., Ernst L.M, & Mestan K.K. (2017). Cord Blood Acute Phase Reactants Predict Early Onset Neonatal Sepsis in Preterm Infants. PLoS ONE, 12(1), e0168677.

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Profiling Soluble Immune Checkpoints in Serum

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Peripheral blood samples were collected at baseline (T0) before the first administration of any treatment. After centrifugation, serum samples were collected and stored at −80 °C until use. The soluble immune checkpoints (sICs) were evaluated through a multiplex assay using the Human Immuno-Oncology Checkpoint 14-plex ProcartaPlex Panel 1 (catalog number EPX14A-15803-901) (Thermo Fischer Scientific, Waltham, MA, USA). We evaluated the following sICs: CTLA4, CD137, PD-L1, PD-L2, PD-1, LAG3, BTLA, IDO, TIM3, CD80, GITR, HVEM, CD27 and CD28. For each patient, 50 μL of serum was used and added to a 96-well plate together with a mixture of magnetic beads coated with antibody according to the manufacturer’s instructions. A biotinylated detection antibody was added to the plate and then bound to phycoerythrin (PE)-conjugated streptavidin. Samples were analyzed using Luminex 200 platform (BioPlex, Bio-Rad, Hercules, CA, USA). Data are expressed in pg/mL and were analyzed with Bio-Plex Manager Software.

Botticelli A., Zizzari I.G., Scagnoli S., Pomati G., Strigari L., Cirillo A., Cerbelli B., Di Filippo A., Napoletano C., Scirocchi F., Rughetti A., Nuti M., Mezi S, & Marchetti P. (2021). The Role of Soluble LAG3 and Soluble Immune Checkpoints Profile in Advanced Head and Neck Cancer: A Pilot Study. Journal of Personalized Medicine, 11(7), 651.

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Maternal Biomarkers in Term and Preterm Birth

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Blood samples of labouring women (either term or preterm) were collected by the attending midwife upon admission to the labour and delivery ward using serum separating tubes. Women at term not in labour were sampled prior to their Caesarean section. Gestational age matched controls were enrolled from the prenatal clinic. These pregnant women were screened at 20-22 weeks (structural ultrasound) to verify whether they fulfilled the inclusion criteria. These women were matched for week of gestation with a PTB case. Sampling was performed during a subsequent prenatal consultation at the appropriate gestational age.
Samples were stored at 4°C until centrifugation. Blood samples were centrifuged at 1000 g for 10 minutes at room temperature to harvest serum. All samples were stored at -80 °C until analysis.
sTREM-1 concentrations were determined using an enzyme-linked immunoassay. Concentrations of MMP-3, -9 and -13 and TIMP-1, -2, -3 and -4 were assessed using a Human Matrix Metalloproteinases 3-Plex Panel and Human TIMP Multiplex Kit. Multiple proteins were analysed using a multiplex bead immunoassay (Human Cytokine 30-plex Panel). All multiplexes were run on a Luminex 200 platform equipped with Bio-Plex software.

, & Tency I. (2014). Inflammatory response in maternal serum during preterm labour. Facts, Views & Vision in ObGyn, 6(1), 19-30.

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Multiplex Serum Cytokine Profiling

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Serum, collected at baseline (T0), was assayed to detect the concentration of 12 cytokines, 5 chemokines, 13 sICs, 3 adhesion molecules and IDO. Levels of soluble immune related molecules were dosed through a multiplex assay using the ProcartaPlex Human Inflammation Panel (20 Plex, catalog number EPX200-12185-901; sE-Selectin; GM-CSF; ICAM-1/CD54; IFN alpha; IFN gamma; IL-1 alpha; IL-1 beta; IL-4; IL-6; IL-8; IL-10; IL-12p70; IL-13; IL-17A/CTLA-8; IP-10/CXCL10; MCP-1/CCL2; MIP-1alpha/CCL3; MIP-1 beta/CCL4; sP-Selectin; TNF alpha) (eBioscence, Vienna, Austria) and the Human Immu-no-Oncology Checkpoint 14-Plex ProcartaPlex Panel 1 (catalog number EPX14A-15803-901; BTLA; GITR; HVEM; IDO; LAG-3; PD1; PD-L1; PD-L2; TIM-3; CD28; CD80; CD137; CD27; CD152) (eBioscence) according to manufacturer instruction. Samples were measured using Luminex 200 platform (BioPlex, Bio-Rad) and data, expressed in pg/ml of protein, were analyzed using Bio-Plex Manager Software.

Mezi S., Pomati G., Fiscon G., Amirhassankhani S., Zizzari I.G., Napoletano C., Rughetti A., Rossi E., Schinzari G., Tortora G., Lanzetta G., D’Amati G., Nuti M., Santini D, & Botticelli A. (2023). A network approach to define the predictive role of immune profile on tumor response and toxicity of anti PD-1 single agent immunotherapy in patients with solid tumors. Frontiers in Immunology, 14, 1199089.

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Quantifying Cytokines and Checkpoint Molecules in mRCC

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The sera from mRCC patients were assayed to quantify cytokines and soluble checkpoint molecules using the ProcartaPlex Human Inflammation Panel (20 Plex, catalog number EPX200-12185-901; sE-Selectin; GM-CSF; ICAM-1/CD54; IFN alpha; IFN gamma; IL-1 alpha; IL-1 beta; IL-4; IL-6; IL-8; IL-10; IL-12p70; IL-13; IL-17A/CTLA-8; IP-10/CXCL10; MCP-1/CCL2; MIP-1alpha/CCL3; MIP-1 beta/CCL4; sP-Selectin; TNF alpha) (eBioscence, Vienna, Austria) and the Human Immuno-Oncology Checkpoint 14-Plex ProcartaPlex Panel 1 (catalog number EPX14A-15803-901; BTLA; GITR; HVEM; IDO; LAG-3: 47; PD1; PD-L1; PD-L2; TIM-3; CD28; CD80; CD137; CD27; CD152) (eBioscence). Samples were measured using Luminex 200 platform (BioPlex; Bio-Rad, Bio-Rad, Hercules, CA, USA) and data, expressed in pg/mL of protein, were analyzed using Bio-Plex Manager Software (version 6.1, Bio-Rad).

Zizzari I.G., Napoletano C., Di Filippo A., Botticelli A., Gelibter A., Calabrò F., Rossi E., Schinzari G., Urbano F., Pomati G., Scagnoli S., Rughetti A., Caponnetto S., Marchetti P, & Nuti M. (2020). Exploratory Pilot Study of Circulating Biomarkers in Metastatic Renal Cell Carcinoma. Cancers, 12(9), 2620.

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Multiplex Analysis of Inflammatory Markers

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All measurements were made in duplicate using commercially available kits and manufacturers guidelines. Serum IL-1β, IL-4, IL-6, IL-8, IL-10, IL-13, IL-17, GMC-SF, TNFα and MCP-1 were simultaneously measured by multiplex luminometry (Bio-Rad, Hemel Hempstead, UK). Samples were analysed using a Bio-Plex Luminex²⁰⁰ platform equipped with a 635 nm red and 532 nm green laser using Bio-Plex Manager™ software. Detection of C-reactive protein (CRP) was by high sensitivity ELISA using a commercial kit (IBL International, Hamburg, Germany). Plasma adiponectin and leptin were assessed separately by solid phase sandwich ELISA (R&D Systems, Abingdon, UK). Plasma insulin levels were determined by sandwich ELISA (Life Technologies, Paisley, UK). Plasma glucose, cholesterol and triglycerides (Instrumentation Laboratories, Warrington, UK) were determined photometrically on an automated chemistry analyser (iLab 650, Instrumentation Laboratories, Warrington, UK).

Bartlett D.B., Fox O., McNulty C.L., Greenwood H.L., Murphy L., Sapey E., Goodman M., Crabtree N., Thøgersen-Ntoumani C., Fisher J.P., Wagenmakers A.J, & Lord J.M. (2016). Habitual physical activity is associated with the maintenance of neutrophil migratory dynamics in healthy older adults. Brain, Behavior, and Immunity, 56, 12-20.

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Multiplex Immunoassay for Serum Biomarkers

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For each patient, 50 µL of serum was used and added to a 96-well plate, together with a mixture of magnetic beads coated with antibody, according to the manufacturer’s instructions. Serum concentration of cytokines (TNF alpha, IFN alpha, IFN gamma, IL1 alpha, IL1 beta, IL10, IL12p70, IL13, IL17A, IL4, IL6, GM-CSF), chemokines (MCP1, MIP-1alpha, MIP-1 beta, IL8, IP10), soluble immune checkpoints (BTLA, sCD137 sCD27, sCD28, sCD80, sCTLA-4, sGITR, HEVM, sLAG3, sPD1, sPDL-1, sPDL-2, sTIM3), adhesion molecules (sE-selectin, sP-selectin, sI-CAM-1), and IDO were evaluated. The concentration of the molecules was dosed through a multiplex assay using the Human Immuno-Oncology Checkpoint 14-plex ProcartaPlex panel 1 (catalog number: EPX14A-15803-901) (eBioscience) (Thermo Fischer Scientific, Waltham, MA, USA) and the Human Immuno-Oncology Checkpoint 14-plex ProcartaPlex Human Inflammation panel (catalog number: EPX200-12185-901). Samples were analyzed using Luminex 200 platform (BioPlex, Bio-Rad, Hercules, CA, USA). Data (expressed in pg/mL) were analyzed with Bio-Plex Manager Software.

Mezi S., Pomati G., Zizzari I.G., Di Filippo A., Cerbelli B., Cirillo A., Fiscon G., Amirhassankhani S., Valentini V., De Vincentiis M., Corsi A., Di Gioia C., Tombolini V., Della Rocca C., Polimeni A., Nuti M., Marchetti P, & Botticelli A. (2022). Genomic and Immune Approach in Platinum Refractory HPV-Negative Head and Neck Squamous Cell Carcinoma Patients Treated with Immunotherapy: A Novel Combined Profile. Biomedicines, 10(11), 2732.

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Multiplex Cytokine Profiling in Renal Tissue

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Multiplex kits (R&D Systems, Minneapolis, MN) were used on the Luminex 200 platform (with Bioplex software) to determine renal cytokine levels. Renal tissue was collected and homogenized in PBS with protease inhibitors. Analysis of renal interleukin (IL) 1β, IL-2, IL-6, IL-17, IL-10, tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) were performed following the protocol provided by the manufacturer and expressed as renal cytokine per mg of renal protein. Protein levels were quantified by Bradford assay.

Asirvatham-Jeyaraj N., Fiege J.K., Han R., Foss J., Banek C.T., Burbach B.J., Razzoli M., Bartolomucci A., Shimizu Y., Panoskaltsis-Mortari A, & Osborn J.W. (2016). RENAL DENERVATION NORMALIZES ARTERIAL PRESSURE WITH NO EFFECT ON GLUCOSE METABOLISM OR RENAL INFLAMMATION IN OBESE HYPERTENSIVE MICE. Hypertension (Dallas, Tex. : 1979), 68(4), 929-936.

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Quantifying Mouse Serum MMPs

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Levels of matrix metalloproteinases (MMPs) in the serum of mice were measured using a Milliplex Map Human MMP panel 2 kit (R & D system, Minneapolis, MN). The Luminex 200 platform, coupled with BioRad Bio-Plex software (BioRad), was used to measure the MMP levels according to the manufacturer's protocols.

Woo B.H., Kim D.J., Choi J.I., Kim S.J., Park B.S., Song J.M., Lee J.H, & Park H.R. (2017). Oral cancer cells sustainedly infected with Porphyromonas gingivalis exhibit resistance to Taxol and have higher metastatic potential. Oncotarget, 8(29), 46981-46992.

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