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.
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.
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:
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.
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