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CRO Diaries: Using Steroids In MSD-ECL And ELISA

Steroid hormones are crucial for several body functions that range from an inflammatory response to the regulation of metabolic pathways. Generally, the adrenal cortex and gonads synthesize and release steroids in the bloodstream. Structurally, a steroid is a biological compound consisting of four rings arranged in a molecular configuration. They primarily function in two ways, signaling molecules and core cell membrane components.

Today, companies produce steroids in laboratories. These man-made or artificial steroids are of two types, corticosteroids and anabolic-androgenic steroids. Corticosteroids are used to fight inflammation in the body. They work like cortisol, a hormone that controls the immune system from causing inflammation in the body. On the other hand, anabolic-androgenic steroids are a version of testosterone that help build bigger muscles.

These steroids have to go through several phases of drug development.  After passing each stage of the drug discovery and development process, steroids become available to the general public. Hence,  assessing and measuring steroids in the body is well-understood in the medical community. Meso Scale Discovery electrochemiluminescence (MSD-ECL) and ELISA assays are widely used to detect and measure steroids. Therefore, let us individually focus on MSD-ECL and ELISA for evaluating steroids.

Using steroids in MSD-ECL

Mesoscale multiplex assays can detect structurally similar steroids even at low levels. Hormones such as testosterone, estradiol, dehydroepiandrosterone, progesterone, and luteinizing hormone, are highly conserved across different species. MSD multiplex panels can specifically and quantifiably detect these hormones. MSD’s Steroid Hormone Panel 1 is a crucial preclinical assay that can be vital in IND-enabling studies. The panel simultaneously measures testosterone, estradiol, dehydroepiandrosterone, and progesterone in a single sample. Besides, MSD singleplex assay can efficiently detect a luteinizing hormone in urine and blood samples.

MSD ELISA assays use SULFO-TAG labels that, upon electrochemical stimulation, emit lights for analyte detection. Some vital features of MSD-ECL assays are:

  • Higher signal to background ratio and minimal background signals
  • Only analytes near the electrode are detected
  • Non-radioactive, stable, and conveniently conjugated labels
  • Emission at around 620 nm and hence do not have color quenching issues
  • And signal amplification.

Using steroids in ELISA

In practice, ELISA assays are one of the most used enzyme immunoassays. There are several ELISA formats available for analyte detection. However, for steroids and other low molecular weight compounds, indirect competitive ELISA is the most suitable format. High sensitivity and measurements in different biological matrices are characteristic features of ELISA assays. Besides, chemiluminescent enzyme immunoassays are increasingly used in clinical diagnostics. These assays are sensitive and have a dynamic detection range. Most importantly, they have a shorter working protocol than compared to traditional ELISA assays. Additionally, chemiluminescent enzyme immunoassays are also not demanding in terms of instrumentation. 

ELISA assays for steroids are unique for each analyte. Each analyte will need a unique approach for the immobilization strategy, the antibodies, the assay matrix, sensitivity and specificity of detection, and synthesis of immunogens. However, ELISA assays can detect steroids from various matrices of plant, animal, or pharmaceutical origin with detection limits of tens of picograms/ml.

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