How to Choose the Best Biomarker Kit for Your Research

How to Choose the Best Biomarker Kit for Your Research

Whether your research needs a broad cytokine array or the analysis of a unique biomarker, the following content provides an outline of the recommended key parameters that will guide you in finding the best multiplex biomarker assay.

Quantitative range of detection and sensitivity

Before committing to running your precious samples on a particular biomarker kit, review the protocol and specifically check the quantitative range of detection and the sensitivity. It is important to determine whether the assay will provide sufficient sensitivity and range of detection. Please note, however, the range of detection reported by the vendor may not reflect the range of detection that will be encountered in a specific sample type.  Two reasons for this discrepancy:  First, biomarker kit manufacturers base their range of detection on recombinant protein whereas your sample’s results will be based on the detection of native proteins.  Second, the matrix of different samples will vary. If the true quantitative range for native proteins is unknown or questionable, the user should ensure the range of detection for each biomarker is suitable for their samples. This can be accomplished by conducting a pilot run.

The vendors Eve Technologies utilizes typically provide standard curve details (assay detection ranges) for their biomarker multiplex assay kits and often include minimal detectable concentrations for each biomarker. For our routine Discovery Assays, these details are included in their respective webpages.

Performance in sample matrix

Commercially available kits typically provide a protein-based assay buffer and some will even provide an additional clean serum matrix to match a standard curve’s protein content you’re your serum or plasma samples. When biomarker kit manufacturers validate the performance of specific samples they often run a simple spike and recovery analysis where recombinant cytokines or other biomarker proteins are spiked into specific sample types at various concentrations.  As mentioned above, this method is not validating the presence of native detectable target proteins.  Inquire with the biomarker kit manufacturer if their validation includes results showing good detectable levels of actual native protein in your particular sample type.  In cases where the biomarker kit vendor cannot confirm performance with the intended sample type/matrix, carry out a pilot run to determine whether the kit can detect native protein levels in your samples before running all samples or placing a large order for multiple kits.

Vendor support

The biomarker kit market has become flooded with newly available biomarker detection kits along with dozens of new kit manufacturers.  Many of these new kits / vendors are selling poorly validated kits or even kits that simply do not meet minimal performance standards.  Unless the detection of your biomarker is only offered by a single vendor, select a biomarker kit provider that is reputable and has adequate technical support.  Many vendors today are not actually manufacturing the kits they are selling and are merely acting as a distributor.  Because of this, they lack in knowledge of the kits they are selling and have very poor technical support.  For the biomarker kit that you are interested in, ask the vendor where it is made.

With Eve Technologies’ 15 plus years of experience we have built long-standing relationships with our preferred vendors. We are always assessing and evaluating the multiplex kits we provide to our clients to ensure the best results possible. We are always in communication with vendors regarding manufacturing changes such as reagent changes.  If a requested biomarker is not available in a multiplex format from a reputable company we will work on meeting our client’s needs by utilizing a lesser known vendor’s ELISA or colorimetric assay, however this is done with the caveat that we cannot guarantee the results or the kit’s performance.

To summarize, the 3 above criteria should always be considered when choosing the platform and kits for your research. If something happens to be lacking in any of the above, a different technology platform or biomarker kit provider should be considered.

Multiplex Biomarker Kit Platforms

TechnologySurfaceDetectionBenefits & Drawbacks
Luminex / Milliplex / Bio-PlexFluorescent beadsFluorescence-Efficient multiplexing   -Sensitivity
-Vast Biomarker menu
-Custom assay development
-Regulatory path from RUO to -IVD/CDx
-Flexibility to build your own assay -Licensed and reputable suppliers -Matrix interference challenges (solved at Eve Tech)
BioscaleAcoustic membrane particle technologySensor resonant frequency-Sensitivity   -Limited market penetration
QuanterixMagnetic beads (SiMOA)Fluorescence-Ultra High Sensitivity
-Custom assay development
-Regulatory path from RUO to -IVD/CDx-Large footprint & Capital intensive
Flow Cytometry + beadsMagnetic beadsFluorescence-Biomarker menu   -Limited multiplex ability -Limited dynamic range and sensitivity
Meso Scale DiscoveryElectrode carbon surfaceECL-Validation & Sensitivity
-Biomarker menu
-Custom assay development
-Automation-Low multiplex is key drawback- Business model prevents kit purchase if using second-hand equipment
AushonQuartzDCD-Ultra High Sensitivity
-Custom assay development-Limited multiplex and analyte menu
-Custom assay development
-Regulatory path from RUO to -IVD/CDx-Limited kit menu-Not intended for basic research
GenalyteSilicon photonic biosensorsSurface resonance units-Sensitivity & speed
-Elimination of cross-talk-Limited biomarker menu

F L fluorescence, RUO research uses only, IVD/CDx in vitro diagnostic/companion diagnostic, SiMOA single molecule array

We are always here to assist and can be contacted at the below:

Phone: (+01) 403-210-9351 or 403-861-0108 Fax: 403-239-0481



Jani, Darshana, et al. “Recommendations for use and fit-for-purpose validation of biomarker multiplex ligand binding assays in drug development.” The AAPS journal 18.1 (2016): 1-14..