Technology

Multiplexing LASER Bead Technology

General Multiplex Info

The term multiplexing was originally used in electronics in reference to the transmission of multiple signals at the same time. The biotechnical world has adopted this term to describe simultaneous testing of multiple targets in a single assay. Today, a number of companies such as Millipore (Milliplex) and Bio-Rad (Bio-Plex) develop kits on this technology platform which allows for the simultaneous detection of up to 100 different analytes in a single well. Multiplexing LASER Bead Technology is at the forefront of biomedical testing and facilitates studies of numerous analyte categories such as cytokines, chemokines and growth factors. This technology recently received acclamation as one of the most advanced and useful technologies, and was the 2006 recipient of the Frost & Sullivan Technology Innovation & Leadership of the Year Award.

How Multiplexing works

The multiplexing technology that Eve Technologies uses is based on color-coded polystyrene beads. Bead coloration is achieved by utilizing different concentrations of red and infrared fluorophore dyes to create 100 uniquely-colored bead sets. Considering that these bead sets contain a unique color / fluorophore signature (that can be individually identified by the bead analyzer such as a Bio-Plex 200), they can then be combined within the same assay well. For example, a capture antibody for IL-6 is coupled to a population of dark red beads, and a capture antibody for GM-CSF is coupled to a population of pink beads. These two antibody-coupled beads can now be mixed together forming a 2-plex. Each analyte is distinguished from the other because they are bound to differently colored/fluorescent beads.

The bead analyzer (Bio-Plex 200) includes a dual-laser system and a flow-cytometry system. One laser activates the fluorescent dye within the beads which identifies the specific analyte. The second laser excites the fluorescent conjugate (streptavidin-phycoerythrin) that has been bound to the beads during the assay. The amount of the conjugate detected by the analyzer is in direct proportion to the amount of the target analyte. The results are quantified according to a standard curve.

Assay Steps in Detecting Fluorescent Intensities

Multiplex Assay Benefits and Applications

By enabling users to perform up to 100 bioassays simultaneously on a single sample, these assays are efficient, cost effective and have high accuracy ratings. Even single-plex assays (1 analyte target) on this platform are more efficient and cost effective than commonly used assay platforms such as ELISA. Today, there are hundreds of analyte-coupled beads available that can be combined and multiplexed together with other beads. Examples include cytokine arrays, determining the presence or titre of specific antibodies, gene expression profiling, detecting and quantifying components of cellular signaling and detecting the presence of cancer markers.

In addition to being highly efficient, these Multiplexing Laser Bead assays have a remarkable low demand on sample volume – often requiring only 2-25 µl of plasma, serum or other sample types. This multiplexing technology requires much less sample volume than ELISA or Western Blot.

Meso Scale Discovery® (MSD)

General MSD® Info:

In addition to our Multiplexing LASER Bead Technology, Eve Technologies also utilizes an electrochemiluminescence (ECL) technology platform designed by Meso Scale Discovery ® (MSD). Being a premier platform in the field of immunoassays, MSD® technology stands out for its exceptional sensitivity and precision.  These attributes make it a powerful tool for biomedical research, drug discovery, and clinical trials. MSD®’s ability to detect and quantify biomarkers with remarkable accuracy enables research applications that would otherwise be limited due to low-levels of biomarkers, low sample volume, and other challenging variables.

How MSD® Works:

MSD® technology utilizes electrochemiluminescence (ECL) to achieve high sensitivity and precision in detecting analytes. The process involves several key steps:

• Sample Preparation: Samples are prepared according to specific assay protocols to ensure optimal conditions for analyte detection.

• Assay Assembly: Capture antibodies specific to the target analytes are immobilized on the MSD® plates. Samples are then added to these plates, allowing the target analytes to bind to the capture antibodies.

• Signal Generation: Detection antibodies, labeled with electrochemiluminescent tags, are introduced. These tags emit light when excited by an electrical current applied to the plate electrodes.

• Result Analysis: The emitted light is measured by an MSD® instrument. The intensity of the light signal is directly proportional to the concentration of the analyte in the sample, allowing for precise quantification.

MSD® Assay Benefits and Applications:

• Exceptional Sensitivity: : MSD® assays can detect analytes at very low concentrations, often in the femtogram/mL range. This high sensitivity is crucial for identifying low-abundance biomarkers, which are often undetectable with other platforms.

• High Precision: The assays offer minimal background noise and superior signal-to-noise ratios, ensuring reliable and reproducible results even in complex sample matrices. This precision is vital for accurate quantification in research and diagnostic applications.

• Multiplexing Capability: Though not as vast as the Multiplexing LASER Bead Technology platform, the MSD® platform also allows for the simultaneous detection of multiple analytes in a single sample. This capability enhances throughput, reduces sample volume requirements, and conserves valuable samples, making it ideal for comprehensive biomarker profiling.

• Cost-Effectiveness: Through Eve Technologies’ Discovery Assays®, select MSD® assays are designed to be cost-effective by offering flexible purchase models based on the required number of wells. By aligning pricing with Eve Technologies’ mandate, we are ensuring that the MSD® technology platform is accessible to a wide range of research budgets.

• Versatile Applications: MSD® supports a wide range of applications, including biomarker discovery, drug development, therapeutic monitoring, and basic research. Its flexibility and adaptability enable researchers to tailor assays to meet specific research needs, facilitating a broad spectrum of scientific inquiries.  Examples of this are the development of custom anti-drug antibody assays (ADA studies) and pharmacokinetics (PK studies).

• Streamlined Workflow: Though Assays on the MSD® platform are highly complex, the analyzer offers strong efficiencies in analysis and data generation.  When you partner with Eve Technologies, we take care of the assay challenges associated with MSD® and simply provide you with your results within industry-leading turn-around-times.