Analytical Assays

Multiplex analysis

Multiplex immunoassays allow simultaneous detection and quantification of multiple analytes in a single sample from a variety of biological specimens including but not limited to serum, plasma, CSF, cell culture supernatants, sputum.

xMAP Technology has application in serological assays by easy detection of antibodies. A serology assay enables the detection of an antibody (target) with the use of a capture protein antigen attached to the surface of a microsphere. A detection antibody that incorporates a fluorescent label is used to quantify the amount of target antibody present. This assay is useful when serum antibodies need to be measured, such as for the response to a vaccine, allergic reactions, or autoantibody detection.

It combines the efficiency and sensitivity of traditional ELISAs with high through-put capacity, small sample volume,efficiency in terms of time and cost,high reproducibility and sensitivityand analyte quantification over a wide range of concentrations.

Principle of xMAP Technology

LuminexxMAP platform is a robust multiplexing technology which combines advanced fluidics, optics, and digital signal processing with proprietary microsphere technology to deliver multiplexed assay capabilities.The LuminexxMAP (Multi-Analyte Profiling) technology uses 5.5-µm microspheres that are labelled with two fluorescent dyes (red and infrared) at ten different concentrations to produce up to 100 different sets of microspheres. Each set of microspheres can then be derivatized by a specific type of capture ligand. Current multiplexed immunoassays are based on multi-marker strategies, in which high- affinity capture ligands (antibodies or proteins/peptides) are immobilized in parallel assays. Following incubation with biological samples, target analytes are bound to corresponding capture ligands. After washing to remove unbound proteins, captured targets are usually detected by using various labeled reporter ligands. Then target analytes are quantified by measuring the signal intensity of the detection label, which is converted to mass units of target analyte using calibration curves. Luminex MAGPIXTM system which is based on the principle of fluorescence imaging, where Light Emitting Diodes (LEDs) and a CCD camera were used to replace the lasers and Photo Multiplying Tubes (PMTs) to deliver a cost-effective, compact, and reliable multiplexing platform. Apart with Multiplex Immunoassays, cellular assays have always been a strong tool in the research lab. MAGPIX® system immobilizes beads with magnetic force, excites them using light-emitting diodes (LEDs), and detects beads and analytes using a CCD camera. The result is a more compact, robust, and cost-effective multiplexing tool.
Courtesy: www.bio-rad.com

Applications:

  1. Measurement of cytokine/chemokine/inflammatory mediator expression in variety of biological specimens which can be applied for biomarker discovery and Target validation
  2. Allergy testing
  3. Single nucleotide polymorphisms (SNPs) testing
  4. Diagnosis of infectious diseases
  5. Application areas include inflammation, autoimmune diseases, asthma, obesity/diabetes, neurological disorders, cancer, cardiovascular disease, drug discovery and vaccine development.

Biomarker qualification and validation

  • Robust validated technology 
  • Simultaneous detection and quantification of multiple analytes in a single sample from a variety of biological specimens
  • High sensitivity and reproducibility
  • High through-put capacity
  • Efficiency in terms of time and cost
  • Various species

Popular Protein Analytes for Measurement

No Plex Assay Panel Description
1 41-plex Human Cytokine/Chemokine Panel I sCD40L, EGF, Eotaxin/CCL11, FGF-2, Flt-3 ligand, Fractalkine, G-CSF, GM-CSF, GRO, IFN-α2, IFN-γ, IL-1α, IL-1β, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17, IP-10, MCP-1, MCP-3, MDC (CCL22), MIP-1α, MIP-1β, PDGF-AA, PDGF-AB/BB, RANTES, TGF-α, TNF-α, TNF-β, VEGF
2 29-plex Human Cytokine/Chemokine Panel II EGF, G-CSF, GM-CSF, IFN-α2, IFN-γ, IL-1α, IL-1β, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17, IP-10, MCP-1, MIP-1α, MIP-1β, TNF-α, TNF-β, VEGF, Eotaxin/CCL11
3 23-plex Human Cytokine/Chemokine Panel III 6Ckine, BCA-1, CTACK, ENA-78, Eotaxin-2, Eotaxin-3, I-309, IL-16, IL-20, IL-21, IL-23, IL-28A, IL-33, LIF, MCP-2, MCP-4, MIP-1d, SCF, SDF-1A+β, TARC, TPO, TRAIL, TSLP
4 25-plex Human Th17 Panel GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17A, IL-17F, IL-17E/IL-25, IL-21, IL-22, IL-23, IL-27, IL-28A, IL-31, IL-33, MIP-3α/CCL20, TNF-α, TNFβ
5 5-plex Human Diabetes Panel Insulin, Glucagon, Leptin, GLP-1 (Active), C-Peptide
6 11-plex Human Kidney Injury Panel Calbindin, Collagen IV, FABP1, GSTα, GSTπ, IP-10, KIM-1, Osteoactivin, Renin, TFF-3, TIMP-1
7 8-plex Human Src Family Kinase Panel – Phosphoprotein Blk (Tyr389), Fgr (Tyr412), Fyn (Tyr420), Hck (Tyr411), Lck (Tyr394), Lyn (Tyr397), Src (Tyr419), Yes (Tyr421)
8 17-plex Human Angiogenesis/Growth Factor Panel Angiopoietin-2, BMP-9, EGF, Endoglin, Endothelin-1, FGF-1, FGF-2, Follistatin, G-CSF, HB-EGF, HGF, IL-8, Leptin, PLGF, VEGF-A, VEGF-C, VEGF-D
9 7-plex DNA Damage/Genotoxicity Panel ATR (Total), Chk1 (Ser345), Chk2 (Thr68), H2A.X (Ser139), MDM2 (total), p21 (Total), p53 (Ser15)
10 6-plex Human Apolipoprotein Panel Apo AI, Apo AII, Apo B, Apo CII, Apo CIII, Apo E
11 12-plex Human Bone Metabolism Panel ACTH, DKK-1, FGF-23, IL-1β, IL-6, Insulin, Leptin, Osteocalcin, OPN – Osteopontin, Osteoprotegerin, PTH, SOST, TNF-α
12 9-plex Human Cancer/ Metastasis Biomarker Panel DKK-1, GDF-15, NSE, OPG, Osteonectin (SPARC), Periostin, TRAP5, TWEAK, YKL40
13 10-plex Human Cardiovascular Disease (CVD) Panel 1 ADAMTS13, D-Dimer, GDF-15, Myoglobin, sICAM-1, MPO, P-selectin, Lipocalin-2/NGAL, sVCAM-1, SAA.
14 14-plex Human Cardiovascular Disease (Acute Phase) Panel 2 AGP, Haptoglobin, SAP, Adipsin,  Human a2, CRP, Fetuin, Fibrinogen, L-Selectin, PF4, vonWillebrand Factor
15 24-plex Human Circulating Cancer Biomarker Panel I AFP, CEA, CYFRA21-1, FGF-2, HE4, PSA Total, PSA free, HGF, IL-6, IL-8, Leptin, MIF, OPN – Osteopontin, Prolactin, SCF, sFas, sFasL, TGF-α, TNF-α, TRAIL, VEGF, CA125, CA15-3, CA19-9, β-HCG
16 5-plex Human Circulating Cancer Biomarker Panel 2 Antithrombin III, Complement Factor H, Extracellular Matrix Protein 1, Vitamin D Binding Protein, Vitronectin
17 7-plex Human Early Apoptosis Panel Akt (Ser473), BAD (Ser112), Bcl-2 (Ser70), Active Caspase 8 (Asp384), Active Caspase 9 (Asp315), JNK (Thr183/Tyr185), p53 (Ser46)
18 11-plex Human Kidney Injury Panel 1 Calbindin, Collagen IV, FABP1, GSTα, GSTπ, IP-10, KIM-1, Osteoactivin, Renin, TFF-3, TIMP-1
19 7-plex Human Kidney Injury Panel 2 α-1-Microglobulin, Albumin, Clusterin, Cystatin C, EGF, Lipocalin-2/NGAL, Osteopontin (OPN)
21 13-plex Human Metabolic Hormone Panel Amylin (Total), Amylin (Active), C-Peptide, Ghrelin, GIP, GLP-1 (Active), GLP-1 (Total), Glucagon, IL-6, Insulin, Leptin, MCP-1, PP, PYY, TNF-α
20 9-plex Human Liver Protein Panel – Metabolism AFP, ANGPTL3, ANGPTL4, ANGPTL6/AGF, FABP1, FGF-19, FGF-21, FGF-23, HGF
21 13-plex Human Metabolic Hormone Panel Amylin (Total), Amylin (Active), C-Peptide, Ghrelin, GIP, GLP-1 (Active), GLP-1 (Total), Glucagon, IL-6, Insulin, Leptin, MCP-1, PP, PYY, TNF-α
22 14-plex Human Soluble Cytokine Receptor Panel sCD30, sEGFR, sgp130, sIL-1RI, sIL-1RII, sIL-2Rα, sIL-4R, sIL-6R, sRAGE, sTNFRI, sTNFRII, sVEGF-R1, sVEGF-R2, sVEGF-R3
23 7-plex T-Cell Receptor Signaling Panel CD3ε (pan Tyr), CREB (S133), ERK (Thr185/Tyr187), LAT (pan Tyr), Lck (pan Tyr), Syk (pan Tyr), ZAP-70 (pan Tyr)
24 5-plex Human Neurodegenerative Disease Panel Human Neurodegenerative Disease Panel
25 3-plex Human Sepsis Panel FGF-13, Olfactomedin-4 (OLFM-4), MMP-8
26 7-plex Human Skin Immune Response Panel Cortisol, HSA, Fibronectin, LPS, Involucrin, Keratin-6, Keratin-1,10
27 13-plex Human High Sensitivity T Cell Panel GM-CSF, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, TNF-α
28 7-plex Total Mitogenesis RTK Panel EGFR (Total), ErbB2/HER2 (Total), ErbB3/HER3 (Total), ErbB4/HER4 (Total), IR (total), c-Met/HGFR (Total), IGF-1R (Total)
29 9-plex Multi-Pathway Total Cell Signaling Panel Erk/MAPK 1/2 (Total), Akt (Total), STAT3 (Total), JNK (Total), p70 S6 Kinase (Total), NFκB (Total), STAT5A/B (Total), CREB (Total), p38 (Total)
30 5-plex Human Oxidative Stress Magnetic Bead Panel – Cellular Metabolism Multiplex Assay Catalase, PRX2, SOD1, SOD2, TRX1

ELISA

Serum samples from 4 healthy patients were collected at three time points (pre-, 4 and 24 hours post treatment. The IL-17A levels in these samples were analysed using the ELISA platform.

Flow Cytometry

Western Blot

Biomarker Validation and Qualification

Validation is the process of evaluating the reliability, reproducibility and accuracy of the biomarker across a range of experimental conditions. Qualification process provides an evidentiary link between the biomarker and biological processes/clinical end points.

Following validation and qualification, a biomarker can be used to diagnose the risk, presence and prognosis of a disease in an individual. In evaluating potential drug therapies, a biomarker may be used as a surrogate for a natural endpoint such as survival or irreversible morbidity. If a treatment alters the biomarker, which has a direct connection to improved health, the biomarker serves as a surrogate endpoint for evaluating clinical benefit. Some of the main areas in which molecular biomarkers are used in the drug development process are: early drug development studies, safety studies, proof of concept studies, and molecular profiling.

We offer assay development and validation for prognostic/predictive clinical biomarkers and pathway activation biomarkers:

  • Mutliplex immunoassay
  • Multiplex nucleic acid testing
  • Western Blot
  • Clinical Chemistry assays

Get a consultation today

We offer more possibilities to meet your every need.
Cellomatics Biosciences Ltd Logo
Cellomatics Biosciences Ltd. is a specialised Contract Research Organisation (CRO) providing bespoke preclinical laboratory based services.

Contact Details