Drug absorption assessment is crucial in investigating ADME of novel pharmaceuticals. Creative Biolabs provides Caco-2 permeability assay in our portfolio of in vitro drug absorption services to predict human intestinal permeability and to investigate drug efflux.

Caco-2 cell line is derived from human colon epithelial cancer and it is usually used as a model of human intestinal absorption of drugs. When cultured, the cells spontaneously differentiate into monolayers of polarized enterocytes to serve as a paracellular movement model. In addition to determine the permeability by passive diffusion, Caco-2 cells can also assess the permeability by active uptake/efflux and paracellular active transport as they can express transporter proteins, efflux proteins, and Phase II conjugation enzymes. In many cases, the Caco-2 cell monolayer mimics the human intestinal epithelium. The major functional difference between normal intestinal cells and Caco-2 cells is that the Caco-2 cells lack expression of the cytochrome P450 isozymes and in particular, CYP3A4.

Caco-2 Cell Monolayers Model

Caco-2 Permeability Figure 1. Caco-2 intestinal epithelial cell assay for intestinal absorption. (Li et al. 2003)

The Caco-2 cell monolayer permeability assay is reliable, easy to carry out, and requires only small quantities of compounds. A solution of a test compound is placed on either the apical or basolateral side of the monolayer. After incubation for certain length of time (e.g. 2 h), the concentration of test compound and its rate of transport are measured by chromatography-mass spectrometry (LC-MS) or LC-tandem mass spectrometry (LC-MS-MS).

Assessing the permeability of the test compounds on both apical side (apical to basolateral (AP-BL)) and basolateral side (basolateral to apical (BL-AP)) enables to determine whether a compound undergoes active transport/efflux. The P-glycoprotein (P-gp) inhibitor can be included to test whether active transport is mediated by P-gp.

Calculation of Permeability Coefficient

The transport rate for a particular concentration of a test compound is typically expressed as apparent permeability coefficient (Papp), which is calculated using the following equation:

Caco-2 Permeability

Vr: The volume of the recipient compartment
dC/dt: The slope of the cumulative concentration of the compound in the recipient chamber over time
A: The membrane surface area
C0: The initial compound concentration in the donor chamber

High Throughput Caco-2 Cell Monolayer Assays

Creative Biolabs delivers high-quality data with a high throughput automated approach. We use 6-well, 12-well, 24-well or 96-well cell culture plates for the parallel culturing and testing of Caco-2 cell monolayers. The use of multi-well plates improves the efficiency and reduces overall cost of Caco-2 cell monolayer assays.

Creative Biolabs provides high-quality Caco-2 permeability assay to predict human intestinal permeability and to investigate drug efflux. For more detailed information, please feel free to contact us or directly sent us an inquiry.

References

  1. Li Y, Shin YG, Yu C et al.(2003) “Increasing the throughput and productivity of Caco-2 cell permeability assays using liquid chromatography-mass spectrometry: application to resveratrol absorption and metabolism.” Comb. Chem. High Throughput Screen. 6:757-767
  2. van Breemen RB and Li Y (2005). “Caco-2 cell permeability assays to measure drug absorption.” Expert Opin Drug Metab Toxicol 1(2):175-185

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