Use of Baculovirus BacMam vectors for expression of ABC drug transporters in mammalian cells

Monday, October 24, 2011 — Poster Session I
Noon – 2:00 p.m.	Natcher Conference Center	NCI	PHARM/PHYS-2

Authors

• C Schwartz
• K Kapoor
• SV Ambudkar
• S Shukla

Abstract

ABC drug transporters are expressed in many tissues such as the intestine, liver and brain, and play key roles in drug absorption, distribution, and excretion. In this study, we report the use of BacMam (Baculovirus based expression in mammalian cells) expression system to study function of two major ABC drug transporters, Pgp and ABCG2. We show here that BacMam baculovirus transduced cells have a similar level of cell surface expression and function of these transporters as drug-selected resistant cell lines that overexpress them. Transient expression of Pgp was maintained in these cells for up to 72 h after transduction and 48 h after removal of the BacMam virus. The membranes isolated from BacMam transduced mammalian cells were also used to study the activity of these transporters by substrate-binding and ATPase assays. In addition, we show that the BacMam expression system can be used to co-express and study functions of both Pgp and ABCG2 in these cell lines. Collectively data from this study suggests that BacMam based expression of Pgp and ABCG2 in mammalian cells can be developed as an over expression system to study the transport function and biochemical studies of these proteins in the same cell type.

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