Multiplex immunoassay Biomarker Testing

MULTIPLEX ASSAY SERVICES

Looking to get most data out of limited samples? Boster Bio offers multiplex ELISA service at industry's lowest rates, starting from $1.00 per datapoint with our premade panels.

Benefits:

  1. require minimal sample volume, as low as 25 µL per well
  2. fast turnaround
  3. low cost for developing custom panels
  4. can mix and match hundreds of biomarkers

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About Boster Bio's Multiplex assay Services

Boster Bio has been a leading manufacturer of singleplex sandwich ELISA kits since 1993. As a manufacturer of ELISA compatible antibodies, we can easily customize multiplex immunoassay panels with hundreds of multiplex-optimized biomarkers. Compared with ELISA which detects a single analyte at a time, multiplex assays allow for simultaneous detection of multiple targets from the same sample, saving both time and precious samples. Using our pre-made panels significantly lowers the cost per analyte.

Available assay formats

We can perform both microplate-based and bead-based multiplex assays. We will recommend platforms based on panel availability per your project needs.

For microplate, we use the Q-Plex™ platform, a chemiluminescent sandwich ELISA microarray. This assay measures up to 18 analytes using only 25 µL of samples per well. Click here to read more details

For bead assays, our experienced assay developers can work with most platforms. Click here to read more details

What is a multiplex Assay?

Multiplex immunoassays, such as multiplex ELISA (enzyme-linked immunosorbent assay), allow for the quantification of multiple analytes in a single sample. Multi-protein signatures lead to improved insights in disease mechanisms, diagnostics, and the effects of personalized medicine. They are suitable for robust, high throughput, standardized, and affordable analysis of protein targets and biomarkers. Planar assays and microbead-based assays are utilized as capture technologies in multiplex systems.

How To Work With Us?

As easy as 1, 2, 3.

1. Select Your Assay


2. Send Samples


3. Receive Results!


BOSTER MULTIPLEX ASSAY ADVANTAGES

Minimal samples

As low as 25 µL/well.

More data points

Also more robust.

Fast turnaround

Get results in 5-10 days.

Custom panels

Mix-and-matchable

Most affordable

Industry leading cost.

We promise You a concierge sourcing experience

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It is our project concierges' mission to make your project experience as smooth and memorable as possible. They are subject matter experts who are easily accessible around the clock, always happy to help you solve problems, make recommendations and sort through options.

Book a meeting with your project concierge

Pick your biomarkers

Featuring hundreds of pre-validated antibodies you can mix and match.

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α-Enolase FASL IL-4 MIF SARS-CoV-2 S1
Adiponection Ferritin IL-5 MIG SARS-CoV-2 S2
AGP FGF IL-6 MIP-1α Serotypes
Ang-2 FGF21 IL-7 MIP-1β sFAS
BDNF Fibrinogen IL-8 MIP-2 sIL-1R1
Carbamylated Fibrinogen Fractalkine IL-10 MMP1 sIL-1R2
Carbamylated Histone FSH IL-12p40 MMP2 sIL-1R3
CD-14 GCSF IL-12p70 MMP3 sIL-1R4
CD-26 Ghrelin IL-13 MMP7 sIL-6R
CD-163 Glucagon IL-15 MMP9 sTFR
CEA GMCSF IL-16 MMP13 Survivin
Citrullinated Enolase HCGβ IL-17α MPO TARC
Citrullinated Fibrinogen HGF IL-18 NSE TCA-3
Citrullinated Histone HRP2 IL-18BPα P4 Thyroglobulin
Cortisol I-309 IL-21 PAD3 TGFβ
C-Peptide IFABP IL-22 PAD4 TIMP-1
CRP IFNα IL-23 PAI-1 TIMP-2
CTACK IFNβ IL-27 PDG TNFα
CWPS IFNγ IL-33 PDGF-BB TNFβ
CXCL-1 IFNω IP-10 PDGF-AB/BB TNFRI
CXCL-5 IGF-1 LDH-Pan PF-4 TNFRII
CYFRA-21 IL-1α LDH-Pf P-Selectin VCAM
E1G IL-1β LDH-Pv RANTES VEGF
E2 IL-1Rα Leptin RBP4 YKL40
EGF IL-2 MCP-1 Resistin
Eotaxin IL-2Rα MCP-2 S100A9
Eotaxin-3 IL-3 MCP-3 SARS-CoV-2 Nucleocapsid
α-Enolase IL-1β IL-6 KC TARC
Eotaxin IL-2 IL-10 MCP-1 TNFα
GMCSF IL-3 IL-12p70 MDC
IFNγ IL-4 IL-13 MIP-1α
IL-1α IL-5 IL-17 RANTES
IFNγ IL-1β IL-4 IL-10 TNFα
IL-1α IL-2 IL-6 IL-12p70
IL-1β IL-6 IL-8 TNFα

MULITPLEX ELISA VS TRADITIONAL TECHNIQUES

Quantitative technologies compared

Technology Advantages Disadvantages Application
Multiplex ELISA
  • · Customizable
  • · Minimal sample required (≤25µL)
  • · Fast & Efficient – highest throughput
  • · Low cost per datapoint
  • · Reliable results
  • · Requires validated antibody pair
  • · Specialized equipment necessary for quantification
Quantify multiple proteins
Western Blot · High Sensitivity & specificity
  • · High cost
  • · Difficult to automate
  • · Prone to false/subjective results
Detection of single protein
ELISA
  • · High sensitivity & specificity
  • · Simple, easy to perform procedure
  • · High efficiency
  • · High cost antibody preparation
  • · Labor intensive
  • · Prone to false/subjective results
  • · Large sample size required (50-700µL)
Quantify single protein
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Why is multiplex ELISA better?

The more the better.

Mulitplex Immunoassays such as the planer multiplex ELISA or microbead based suspension assays allow for the simultaneous detection of multiple analytes in a single well or reaction. Multiplex immunoassays yield a wealth of information on the roles of multiple proteins and other biomolecules in complex biological processes in a single sample, thereby providing clinicians with insight into the identification and assessment of disease progression.

Immunoassays have been widely used in many important areas of pharmaceutical analysis such as diagnosis of diseases, therapeutic drug monitoring, clinical pharmacokinetic and bioequivalence studies in drug discovery and pharmaceutical industries.

Example report

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About Boster Bio

PhD-level support
concierge like service

Experience
since 1993

40,000+
publications

2,000+
ELISA kits developed

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Multiplex Assay formats explained

Overview of two common multiplex formats

The planar format includes platforms such as Boster’s Q–Plex™ array whereby high-affinity capture ligands are immobilised discretely on a solid phase, 2D support, usually in a microtiter plate format. The immobilised ligands are subsequently exposed to treatment with the sample and probing with detection antibodies labelled with a reporter system. The suspension format includes platforms such as Luminex™, whereby high-affinity capture ligands are immobilized discretely on fluorescently activated plastic microbeads and mixed with the sample in liquid phase. Subsequent addition of detection antibodies labelled with a reporter dye enables high-resolution analysis of specific fluorescent signal via flow cytometric methods.

multiplex-assay-formats-explained

Microplate Q-Plex™ Platform Principle

The Q-Plex™ multiplex assays utilize a sandwich enzyme immunoassay technique for the measurement of multiple targets. These assays use two different antibodies specific for their respective targets. Analyte specific antibodies are immobilized in specific locations on the array. Samples or calibrators are pipetted into wells, and after washing away any unbound protein, a mixture containing biotinylated analyte specific antibodies is added, completing the sandwich for each specific arrayed analyte. After washing away unbound biotinylated antibody, streptavidin-horseradish peroxidase (SHRP) is added, washed, and the amount of SHRP remaining on each location of the array is proportional to the amount of the target initially captured. The amount of conjugated enzyme on each location of the array is measured with the addition of a chemiluminescent substrate using a Q-Plex plate reader.

Q-plex benefits:

  • Customizable
  • Premade popular panels available
  • Faster turnaround
  • Lower cost on average

Microbead-based Assay Platform Principle

Microbead-based assays such as the Luminex xMAP technology, utilize the same sandwich ELISA principle as above to multiplex up to 100 different assays within a single sample. This technique involves distinctly colored bead sets, antibody pairs to specific analytes and reporter dyes. One antibody to a specific analyte is attached to a set of beads with the same color, and the second antibody to the analyte is attached to a fluorescent reporter dye label. The use of different colored beads enables the simultaneous multiplex detection of many other analytes in the same sample. A dual detection flow cytometer or Luminex machine is used to sort out the different assays by bead colors in one channel and determine the analyte concentration by measuring the reporter dye fluorescence in another channel.

The advantage of using this platform is that it gives you the ability for large scale screening of many analytes against your sample at one time. However one disadvantage of larger panels may be the loss of resolution for lower expressing markers

Microbead-Based Assay benefits:

  • Customizable for large scale screening
  • Premade popular panels available
  • Faster turnaround

FAQs

Yes. The microbead-based multiplex ELISA technology is more sensitive than traditional, singleplex ELISA, with limits of detection improved by ~10-3 µg/L.

Boster’s multiplex ELISA typically requires ~25 µL of sample per well. In general we require minimum 200 µL per sample. For customized assays, contact our experts for specific volume requirements.

Please contact your Boster Bio service Concierge and he/she will answer any questions you may have and make specialized suggestions. If a biomarker is not available in our pre-validated multiplex panels, we could validate and integrate your biomarker (for diagnostic projects) or test it in singleplex ELISA. Check out our popular ELISA sample testing service at this link.

Please check out our guidelines on how to prepare and label samples before shipping them over to us.

5. What sample types can I use with Boster’s kits?

We can process any and all samples in theory and in most practical cases. We have dealt with plasma or serum samples, cell culture samples, lysate samples, urine, CSF to name a few. If you have samples that you think might cause complications, please contact our project concierges for help.

It depends on the biomarkers of interests. The most common situation where a substance would interfere with an immuneassay is that the substance might chelate the protein of interest and form a comple, shielding the epitope detected by the antibody thus blocking the reaction. The typical substances that bear such risks are anti-coagulents, such as Heparin, EDTA, citrate acid, SDS, used in collecting blood. Each antibody pair/ELISA kit typically informs you which anti-coagulent(s) to avoid. If you have any biomarkers in question feel free to ask us any time. If it is a custom assay you are developing with us, we will figure this out for you.

This information varies depending on the multiplex assay. For each analyte we have validated, there is a general detection range and we will supply details on this once the assay has been determined. Contact our Project Concierge for more information.

We have validated analytes against many target species including human, mouse, rat, porcine, canine, feline, bovine. Please ask us for more details on available analytes and other species via our Project Concierge.

This will be very dependant on your assay requirements, time and budget. We highly recommend talking to our Project Concierge who will work out the best option for you.

Biological samples such as plasma, serum, tissue homogenates, etc. contain many components, such as carbohydrates, proteins, and phospholipids that can interfere with the ability of the antibody pairs to bind to their target. This phenomenon is known as the "Matrix Effect" and is usually displayed as lower OD readings than expected. There are a number of ways to combat this effect including diluting your sample and lowering the amount of analytes tested in each assay. Our Project Concierge will work with you to provide the best solution

Yes, tissue homogenate can be used with the Muliplex assays.

Lower expressing analytes can be difficult to detect particularly in a multiplex assay. So long as they fall within the quantifiable range of Boster’s multiplex assay these will be detected and reported, however if their levels fall outside of this range, we cannot guarantee detection.

Direct comparisons between ELISA and Multiplex Assays for particular analytes can be difficult. The antibodies and standards used in single analyte ELISA assays tend to be different to those used in most Multiplex Assays, and additional complexities due to the matrix effect in multiplex assays also needs to be considered. Correlations and trends can be compared between the two platforms but direct comparisons against absolute figures should be treated with caution.

The Boster Q-Plex Multiplex ELISA assay has <15% CV between kit lots. Samples are usually linear out to a 1 to 64 dilution.

The most common assays would be profiling for cytokines, chemokines, and growth factors. The following is a list of all our assay categories:

  • • Acute Phase Proteins
  • • Adipokine and Adipocyte Biomarker
  • • Allergy Testing
  • • Angiogenesis Biomarker
  • • Apoptosis
  • • Autoimmune/Immunology
  • • Bone Metabolism
  • • Cancer Profiling
  • • Cardiovascular Disease
  • • Cellular Signaling
  • • Cytokines, Chemokines and Growth Factors
  • • Diabetes and Obesity Biomarkers
  • • Endocrine
  • • Gut Hormone
  • • Human Reproductive Hormones
  • • Isotyping
  • • Metabolic Markers
  • • Matrix Metallo-Proteinases
  • • Neuroscience
  • • Skin Biomarkers
  • • Toxicity
  • • Transcription Factors

We typically recommend duplicate testing; however the answer to this question will depend on the purpose of your assay. For publication purposes, duplicate and triplicate testing are the most common. Testing in duplicate or triplicate gives you more assurance that your concentrations are accurate. If, however, you are merely running small pilot studies, testing in single may be suitable and may be more cost effective.

No. This is one of the greatest attributes of Boster’s Multiplex Assay Service as we will accommodate one sample or thousands. Custom-Plex Assays also do not have sample limitations; however it is the most economical to fully use entire 96-well plates. Fully using a 96-well plate typically means increments of 40 samples tested in duplicate (80 wells). 16 wells are typically used for the standard curve.

Yes, the array uses a standard curve to accurately quantify samples and the curve has been designed to encompass the biological ranges for individual proteins.