Enzymatic assay

Efficacy type: Vmax slope (OD/min)
Maximum ACh hydrolyzation
Stock solution: 20 mM in DMSO
Compound quantity < 1 mg

In Brief:

Acetylcholinesterase (AChE) is a serine hydrolase mainly found at neuromuscular junctions and cholinergic brain synapses.

AChE hydrolyzes choline esters. Its principal biological role is termination of impulse transmission at cholinergic synapses, by rapid hydrolysis of the neurotransmitter acetylcholine to acetate and choline. Organophosphorus poisons form covalent bonds with a serine residue at the active site of AChE, and are thus potent irreversible inhibitors of AChE. AChE inhibitors prevent cholinesterase from breaking down acetylcholine, increasing both the level and duration of the neurotransmitter action. The assay is based on the Ellman method in which thiocholine, produced by AChE, reacts with 5,5’-dithiobis (2-nitrobenzoic) acid (DTNB) to form a colorimetric (412 nm) product, the5-thio-2-nitrobenzoate (TNB), proportional to the AChE activity present.

Definitions:

To ensure flawless communication, the following definition section introduces the vocabulary used at INVENesis. You will find this vocabulary on our quotes, raw data files, processed data files, preliminary and final reports. Open the Lexicon below.

Assay Preparation:

  • Test plate: microplate in which an assay is run

  • Mother plate: microplate containing stock solutions
  • Stock solution: solution contained in mother plates out of which dilutions will be made
  • Daugher plate: microplate containing solutions used to treat test plates, usually dilution series

Type of Assays:

  • Organism-based assays : functional assay measuring a phenotypic response on a whole organism. Typically, motility, development, deterrency, repellency.
  • Electrophysiology assays: functional assay measuring the electrical respons of a receptor or an organ to a stimulus
  • Stock solution: solution contained in mother plates out of which dilutions will be made
  • Daugher plate: microplate containing solutions used to treat test plates, usually dilution series

Treatment exposure modalities:

Treatment modalities relate to the way the treatment is applied. Different treatment modalities help define how an active ingredient is most susceptible to act in a field case. Some compounds are more active when eaten, others are more active when applied externally on an organism. The following vocabulary is used in the list of assays:

Tarsal:

  • Tarsal: individuals are allowed to walk on surfaces treated with a formulated active ingredient.
  • Tarsal with choice: only a proportion of the surface accessible to individuals is treated.
  • Tarsal without choice: the whole surface accessible to individuals is treated so that individuals are unable to rest on untreated surfaces.

Topical:

  • Topical: direct application of a known amount of formulated active ingredient on a single individual

Oral:

  • Oral: voluntary feeding on medium treated with a formulated active ingredient.
  • Oral with choice: both a treated and an untreated medium are provided.
  • Oral without choice: only one treated medium is provided.

Immersion:

  • Immersion: individuals are immersed in a treated formulation.
  • Permanent Immersion: individuals are immersed for the whole duration of the assay.
  • Temporary Immersion: the treatment is removed after temporary immersion of individuals.