PAMPA Information Leaflet

PAMPA (parallel artificial membrane permeability assay) is an in vitro assay used to evaluate passive diffusion. 

Pion’s PAMPA kit technology uses a membrane coated with lipid to mimic gastrointestinal conditions and allows the evaluation of the passive permeability of a compound at a range of pH values.

Test compound in buffer is filtered prior to adding into the donor plate of a PAMPA sandwich.  The membrane of an acceptor plate is painted with lipid and acceptor sink buffer added to the wells.  The assembled sandwich is left for 16 h at room temperature in a humidified vessel.  Sampled buffers (T=0, T=16 h) are analysed against a matrix-matched calibration curve by LC-MS and permeability calculated using Pion’s PAMPA Explorer software. 

Standard Assay Conditions

  • 50 µM/10 µM incubation, 0.5% DMSO
  • Up to 3 buffer pHs (5.0, 6.2 and 7.4)
  • N=3 replicates
  • Permeability controls included
  • 16 hour incubation
  • Analysis by LC-MS
PAMPA_Results graph

Figure 1: Compounds of varying permeability  (3 pHs).  N=3 experiments each with 3 replicates. Error bars denote standard deviation.

PAMPA_calibration curve

Figure 2: Example calibration curve.

Deliverables

Permeability in cm/s and assay recovery.

Compound Requirements

20 µL of a 10 mM stock per pH or 1 mg of solid (accurately weighed to 2 decimal places) 

Download Technical Resource Here

In Vitro Assays

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