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External / Inter-laboratory Comparison

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Participation in an, in-vitro/ex-vivo, Interlaboratory Assay Comparison, relating to the assessment of nanomaterial-biological interactions

The EU science hub defines interlaboratory comparisons in the following way: “Interlaboratory comparisons (ILCs) are organised either to check the ability of laboratories to deliver accurate testing results to their customers or to find out whether a certain method performs well and is fit for its intended purposes. The former is usually termed ‘proficiency testing’ and the latter ‘collaborative method validation study’. Only those methods passing the stringent requirements of such a study can be subject to standardisation by European or international standardisation bodies (European Committee for Standardization, CEN, and International Organization for Standardization, ISO).”

As part of the REFINE project, we have selected three in vitro/ex vivo assays for a collaborative method validation study in our drive to progress the field of nanobiomaterials.

Detecting DNA damage with the comet assay

DNA damage can be manifold ranging from DNA strand breaks to chromosomal aberrations or whole chromosome loss. Resulting mutations or genetic instability may subsequently cause different diseases including cancer or may even lead to death. Assessing DNA breakage is thus of significant interest. One of the most common methods to detect DNA strand breaks is the alkaline single cell gel electrophoresis or comet assay. The comet assay has achieved the status of a standard test, in the cascade of assays used to assess the safety of novel pharmaceuticals, nanomaterials (NM) and other chemicals.

NLRP3 inflammasome activation by nanomedicinal products

Inflammasomes are large intracellular multiprotein complexes that can respond to exogenous and endogenous danger signals and control the activation of caspase-1. Several types of inflammasomes are identified, each one having its unique activators and proteins that constitute the inflammasome. The NLRP3 inflammasome is the best characterized inflammasome. Sustained NLRP3 inflammasome activation may result in chronic inflammation and subsequent tissue damage. This sustained activation is implicated in a range of chronic diseases such obesity and type 2 diabetes and Alzheimer’s disease. The assay for inflammasome activation described in the in-vitro assay is based on concomitant measurement of cell viability and IL-β and IL-18 production.

Macrophage polarization to M1 & M2 phenotype for cytokine secretion

Macrophages form an important part of the mononuclear phagocyte system and are known for their ability, and propensity, to engulf nanoparticles and remove them from the body. Given their involvement in immunological responses, and their known interactions with nanoparticles, it is vital to gain an understanding of how nanoparticulate systems may impact on their function. This assay seeks to standardise the, ex vivo, differentiation of monocytes to either M1 or M2 phenotypes prior to the determination of cytokine secretion from cells exposed to nanoparticles. This assay provides well defined phenotypic characteristics of these cells for subsequent experimentation or assessment.

Your Participation to REFINE Project

Inter-laboratory Comparison

As part of the development of these assays, we are organising and interlaboratory comparison to interested laboratories, which are not part of the REFINE consortium. Participation in the ILC, provides a number of advantages to your institution including, but not limited to; familiarisation for your staff and laboratory with potential new in vitro/ex vivo assay standards at an early stage, in depth information and support on the assays performance characteristics and direct access to the specialists developing the assay.

As the aim of this ILC is not just to determine the application of these assays, we also intend to publish the results obtained to inform the relevant stakeholders in the community. If you decide to take part, then you can become one of the co-authors in the publication describing the outcome of the interlaboratory comparison, which builds the basis for the further standardisation process.

SOPs

You will be provided with the test samples, SOPs, description of the work to be performed and the reporting form; All of this is free of charge. You have to cover the cost for performing the assays themselves. It is vital that all work performed is done so according to the SOP and work description.

After completion of the experimental work, results will be sent to the REFINE consortium, using the reporting form for analysis and evaluation. Following this stage, a review meeting will be organised, which present the opportunity to learn more about the different assays performance characteristics. We will provide you with a certificate, that you have participated at the selected interlaboratory comparison.

PLEASE NOTE

Due to the current Covid-19 situation a definitive schedule can not be provided. We will resume the Interlaboratory Assay Comparison once the situation improves

If you are interested and would like to receive more detailed information, then please fill the form below with your details and send it to us.







    We will keep your data only for this interlaboratory comparisons and nothing else. Afterwards all records will be deleted.

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