MICROPLATE READERS
The PHERAstar FSX with its advanced assay stability function enables temperature control between 18o
C and 45o C ensuring stable measuring conditions
Innovations in
cannabinoid research I
Barry Whyte from BMG Labtech discusses how microplate readers could help accelerate drug discovery and development
nterest in cannabinoid research is growing as scientists look for new ways to advance drug discovery and development for
different diseases. This field of study holds promise for new treatments for cancer, neurological disorders, pain management and other conditions. In this context, researchers need new approaches to investigate modes of action, ensure the safety of cannabinoid substances, and help develop novel drugs with reduced side effects. Microplate readers can support a
wide variety of analyses associated with cannabinoid research. The use of high-throughput methods offers time savings, reduced costs, and can help meet the growing demand for scientific analyses. Here, we look at some of the ways microplate readers can provide added value in cannabinoid research.
WHAT MICROPLATE READERS BRING TO CANNABINOID RESEARCH Cannabinoids are a diverse group of naturally occurring or
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synthetic chemical compounds that fall into three main categories: Endocannabinoids are produced in the human body and are responsible for the regulation and control of functions like learning and memory; phytocannabinoids are plant-derived compounds but can partly bind to the same receptors; and synthetic cannabinoid receptor agonists (SCRAs) are made in the laboratory. Microplate readers are often used
to detect phytocannabinoids or their metabolites in medical samples or in forensics. For this purpose, immunoassays such as ELISAs serve as methods for quantification. Chemotyping of plants may also help identify the preferred botanical extracts for use or identify cannabis brought across geographic regions illegally. [1,2] Researchers want to understand
the pharmacology of naturally occurring and synthetic cannabinoids at the receptor level to aid drug discovery. Microplate-based methods using fluorescence polarisation, bioluminescence resonance energy transfer (BRET) or time-resolved
fluorescence resonance energy transfer (TR-FRET) enable the sensitive and reliable detection of receptor binding as well as the identification of downstream signal cascades.
HIGH SENSITIVITY AND RELIABILITY In the following example, TR-FRET was used to study the differential binding of natural cannabinoids to the cannabinoid receptor 1 (CB1). The fluorescent molecule CELT-335 acts as a ligand for CB1. CB1 was additionally labelled with terbium to detect the interaction. When the fluorescent agonist binds to the receptor, energy is transferred from terbium to CELT-335 which consequently emits light at 665 nm in addition to 620 nm derived from the terbium (figure 1A). In competition with 100 nM CELT-335, the binding affinity of 0-10 µM of the phytocannabinoids Δ9-THC, Δ9-THCA and Δ9-THCV was monitored. The TR-FRET ratio was determined using BMG LABTECH’s PHERAstar FSX. The binding affinities were similar
for ∆9-THC and ∆9-THCV with pKi values of 7.2 ± 0.6 and 7.0 ± 0.3,
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