BacMam recombinant baculovirus in transporter expression: a study of BCRP and OATP1B1 |
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Authors: | Hassan Namir J Pountney David J Ellis Catherine Mossakowska Danuta E |
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Institution: | Department of Gene Expression and Protein Biochemistry, GlaxoSmithKline, Harlow, UK. namir.j.hassan@gsk.com |
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Abstract: | Human BCRP and OATP1B1 have recently been identified as important transporters in the absorption, distribution, and elimination of clinically significant drugs. In this report, we illustrate the use of modified baculoviruses, termed BacMam viruses for the expression of functional BCRP and OATP1B1 in mammalian cells. We show a variety of host cells efficiently transduced to express BCRP including HEK 293, LLC-PK, and U-2 OS, where protein levels on the cell-surface were modulated by titrating different amounts of viral inoculum. In addition, using the BODIPY-prazosin efflux assay and the BacMam reagent we illustrate inhibition of BCRP activity with GF120918 or Fumitremorgin C. Furthermore, we present data demonstrating simultaneous expression of BCRP and OATP1B1 in BacMam transduced mammalian cells by simply adding viral inoculum of each transporter. Thus these results indicate that BacMam mediated gene delivery provides a novel and efficient research tool for the investigation of single or multiple transporters in vitro. |
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Keywords: | BCRP ABCG2 MXR Efflux transporter OATP1B1 OATP-C OATP2 SLC21A6 Influx transporter BacMam ADME Pharmacokinetics |
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