Quantitative Immunoassays: SIMOA, Erenna, MSD, Single Analyte & Multiplex Screening

PBL can expedite your R&D work as our technologies and services can be tailored to meet your specific requirements under fit-for-purpose guidelines. For many of your sample testing and screening needs, PBL can help you measure your analyte(s) of interest. From state-of-the-art technologies quantifying into the femtogram/milliliter level to simultaneous detection of up to 16 analytes in a single well, PBL offers a multitude of options to fit your project needs.

 

We understand the value of your experimental samples and execute assays just as you would - consuming a minimum of sample while generating robust results. We can take on work you don't have the capacity to handle yourself. Let us help you develop, customize, or just execute assays on your samples in a timely, professional manner.

 

 

Services include:

 

 

* To view an example of our sample testing report, click here.

Interested in our services?

Quanterix Simoa®

High precision. Unprecedented sensitivity. Digital readout. Take your detection to the next level.

 

  • Single Molecule Array technology delivers femtogram per milliliter (fg/ml) level sensitivity for measurement of low abundance biomarkers in serum, plasma, and other matrices
  • Unique technology provides robust detection for single analytes and select multiplex panel options
  • High precision service offers accurate and reproducible data with < 10% inter- and intra-assay CVs
  • Over 50 analytes available including IL-17A, TNF-α, IL-23, IP-10, TRAIL, IL-4, IL-6, IL-8, IL-10, IL-12p70, Aβ-40, Aβ-42, Tau, and NF-LIGHT®

The unique immunoassay technology provided by the Simoa® platform allows PBL to deliver fg/ml level sensitivity for low abundance biomarkers in serum, plasma, and other matrices. Single Molecule Array technology traps a single bead in a femtoliter-sized well allowing for “digital” measurement of each bead. This approach can increase assay sensitivities several orders of magnitude over conventional bioassays.

 

The combination of this instrument’s markedly heightened sensitivity and its' capacity for digital analyte measurement ensures accuracy and precision across a wide dynamic range and minimizes sample volume requirements.

 

Click here for more information on PBL's Ultra-Sensitive fg/ml Biomarker Quantification Services.

 

 

Figure 1. Quanterix Simoa® Technology Overview

Quanterix Simoa Technology Overview

Unprecedented precision can be obtained using Simoa's "digital" readings of single molecular events. In the image above, individual paramagnetic beads are trapped in femtoliter-sized wells, then sealed over and quantified by SIMOA image analysis.

EMD Millipore (Singulex) Erenna®

Accurate and robust measurement. Increase your understanding. Detect at unprecedented levels.

 

  • Cytokine analysis utilizing Single Molecule Counting (SMC) technology enables sub-pg/ml sensitivity for accurate measurement of low abundance biomarkers in normal or disease sera/plasma
  • Minimal sample dilution requirement with 4-Log+ dynamic range
  • High precision cytokine immunoassays provide reproducible data
  • Extensive analyte menu of over 40 different cytokine assays covering several disease areas including IFN-γ, IL-1a, IL-1b, IL-4, IL-6, IL-8, IL-10, IL-13, IL-15, IL-17A, IL-17F, IL-17 heterodimer, IL-23, GLP-1, and TNF-α

The ability to accurately measure low-abundance analytes present in complex matrices is essential for the profiling and characterization of cytokines and other biomarker analytes. Employing proprietary single molecule counting (SMC) technology with robust microparticle-based Erenna® immunoassays, PBL’s cytokine detection services can provide scientists with sub-pg/ml level measurements of low-abundance analytes in healthy or disease sera and plasma.

 

An increased understanding of the role and regulation of cytokines in disease states results from greater profiling and characterization of their activities in biological responses. Singulex technology can be instrumental in furthering our collective understanding of low-abundance cytokines in complex matrices.

 

Click here for more information on PBL's Ultra-Sensitive fg/ml Cytokine Testing Services.

Figure 2. EMD Millipore (Singulex) Erenna® Technology Overview

Singulex Erenna Technology Overview

Low abundance biomarkers in complex biological samples are transferred to a 96-well plate and incubated with capture antibody-coated microparticles. After washing and incubating with dye-labeled detection antibodes, wells are again washed to remove unbound detection antibodies. An elution buffer is added, and the eluate drawn into the instrument for single-molecule counting.

Meso Scale Discovery® Electrochemiluminescence (MSD-ECL)

Broad dynamic range. Minimal background. High sensitivity. Enhanced signal capture achieved through luminescent detection.

 

  • Ability to target low analyte levels in small sample volumes
  • Robust matrix tolerance including serum, plasma, and TCM, as well as various complex biological matrices
  • Measure high and low abundance analytes (normal and elevated levels) within the same sample with 5-Log+ dynamic range
  • Over 180 validated fit-for-purpose singleplex and preconfigured multiplex panel ELISAs available with excellent performance specifications and lot-to-lot consistency

PBL offers high precision protein quantification services for biomarker detection in a broad range of sample matrices for Human, Non-Human Primate (NHP), Mouse, Rat, and Canine samples. PBL utilizes MSD's validated assay kits (V-Plex) to quantify up to 54 analytes within a single sample. This enables simultaneous comparison of the expression of pertinent cytokines, chemokines, and other biomarkers in samples from diseased and normal patients at different intervals.

 

Multiplex immunoassays on the MSD platform maintain the sensitivity and performance offered by singleplex ELISAs while providing additional benefits such as cost-savings and targeting of several related analytes in a single sample. The ability to quantify compatible analytes while requiring no more than 25 μl of neat sample ensures efficient use of precious sample.

 

Click here for more information on PBL's MSD-ECL Protein Detection Services.

 

 

Figure 3. MSD-ECL Technology Overview

Meso Scale Discovery ECL Technology Overview

Unique detection technology utilizes SULFO-TAGTM labels which emit light upon electrochemical stimulation. The intensity of light generated is captured via CCD camera and pixel intensities quantified. This system provides the basis for achieving measurement of a several log range of biomarker expression levels in a variety of matrices.

Single Analyte ELISA Service

Sensitive detection and robust results. Dedicated, flexible workforce. Reliable, high-quality outcomes.

 

Take advantage of PBL ELISAs or other commercial ELISAs for a wide range of additional analytes using PBL's services.

 

PBL ELISAs:

  • High sensitivity ELISAs available for a comprehensive range of species and analytes
  • Compatible with a variety of matrices including serum, plasma, tissue culture media (TCM), and cell lysates
  • Matrix tolerance for autoimmune disease sera*
  • Reproducible results with low intra- and inter-assay CVs

PBL ELISA kits are heterogeneous antibody sandwich immunoassays. The antigen is captured by an antibody immobilized to the wells in a microtiter plate and a secondary antibody is used to complete the sandwich. The detection system which utilizes horseradish peroxidase (HRP) and tetramethyl-benzidine (TMB) as the enzyme-substrate provides a sensitive and robust assay for the detection and quantification of interferons and cytokines.

 

PBL routinely runs ELISA studies for our clients for the detection and differentiation of interferons and cytokines in various sample matrices. We offer a range of heterogeneous immunoassay services for:

  • Human IFN-Alpha*, Beta*, Gamma, Lambda 1/2/3, Omega, Gamma Receptor 1
  • Mouse IFN-Alpha, Beta, Lambda 2/3
  • Cynomolgus/Rhesus IFN-Alpha, Beta
  • Porcine IFN-Alpha

 

For more information on PBL's Single Analyte Service, click here.

 

Multiplex Screening Service

High sensitivity & specificity using low sample volumes. Cost-effective quantitation of multiple analytes.

 

  • Variety of chemokine and cytokine panels already available for human and mouse targets
  • Unique array of analytes including Type I, II & III IFNs, and other key cytokines crucial to innate and adaptive immunity
  • Compatible with autoimmune sera, plasma, and tissue culture media (TCM)
  • Sample volumes of only 50 μl required to quantify all analytes in a single assay
  • Reproducible results with < 15% intra- and inter-assay CVs for most analytes in difficult matrices

PBL's cost-effective multiplex ELISA services enable simultaneous detection of up to 16 different analytes in a single sample.

 

These ELISAs provide chemiluminescent results on each analyte and allow for a global understanding of ongoing immune responses in diseases ranging from cancer to autoimmunity. Evaluation of one given inflammatory molecule in the context of several others, repeated measurements of the same cytokine panels in the same subjects under different treatment assay conditions, and reliable detection of different proteins across a broad range of concentrations are just some of the benefits provided by this multiplex service.

 

For more information on PBL's Multiplex Screening Service, click here. 

Figure 4. Multiplex Technology Overview

Multiplex Analyte Technology Overview

PBL's multiplex ELISAs employ a technology in which distinct capture antibodies are individually spotted into each well of a 96-well ELISA plate in a defined array. These plates are run similar to a normal ELISA run. An image of the plate is then taken using a CCD camera.

Using image analysis software to perform subsequent analysis, the chemiluminescent results for each analyte allow for a deeper understanding of the cytokine profiles of the samples.

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