Efflux Transporter Substrate Identification assay (Caco-2 cells)

Aim: To determine if a test compound is substrate of an efflux transporter, either P-gp (P-glycoprotein) or BCRP (breast cancer resistance protein multidrug transporter), using the Caco-2 cell model.

Background information: Caco-2 cells are a human colon adenocarcinoma cell line which express efflux transporters, such as P-gp and BCRP. Efflux transporters are involved in drug absorption and disposition. As transporter substrates, drugs can be victims of drug-drug interactions in the clinical practice.

Transport assays are conducted both in the absorption (apical-to-basolateral) and efflux (basolateral-to-apical) directions. Test compound is added to either the apical or basolateral sides of the monolayer in the absence and presence of transporter reference inhibitor. Concentrations are quantified by UPLC-MS/MS using a calibration curve.

Compound requirement: 50 µl 10 mM in DMSO

Turnaround time: 4 weeks

Experimental design:

  • Test compound concentration: 10 µM
  • 24-well plate
  • Number of replicates: 2
  • DMSO concentration: 1%
  • Incubation time: 120 min
  • Probe substrate: nitrofurantoin (BCRP) and colchicine (P-gp)
  • Reference inhibitor: Ko143 (BCRP) and verapamil (P-gp)
  • Monolayer integrity marker: lucifer yellow
  • Analytical method: UPLC-MS/MS with calibration curves

Delivered results:

  • Absorption and efflux permeability (+/- transporter inhibitor)
  • Absorption and efflux recovery (+/- transporter inhibitor)
  • Efflux ratio of the probe substrate (+/- transporter inhibitor)

Dear colleagues, collaborators, and friends,

After more than 30 years of shared endeavours, in which we have grown personally and professionally, and with our emotions on the surface, we announce the closure of our company. We have travelled together through ups and downs, celebrated successes and overcome challenges as a team.

We extend our deepest gratitude to each of you for your dedication, passion, and commitment to a shared goal: discovering new treatments for the benefit and well-being of patients.

As we bid farewell, let’s cherish the memories we’ve created, the bonds we’ve formed, and the impact we’ve made together. Though our paths may diverge, the spirit of collaboration and camaraderie that defines our community will always endure.

We leave proud of the work accomplished and thank you for being an integral part of our journey, wishing you success in all your future challenges.

With heartfelt appreciation,

Welab’s team

We are proud to announce that Welab is partner of this new EU funded project.

In collaboration with 12 partners from 6 countries, Welab Barcelona is establishing the Antivirus Pandemic Preparedness EuropeAn pLatform (APPEAL), a European research initiative aimed at enhancing preparedness for future pandemics. This EU funded collaboration will establish a comprehensive program for the development of broad-spectrum antiviral drugs within a five year time frame ensuring drug affordability and accessibility to low income countries.
Link to the press release: https://lnkd.in/d-jmV2CS