A mass spectrometry based functional assay for the quantitative assessment of ABC transporter activity.
ATP-Binding Cassette (ABC) transporters are highly expressed in pharmacological barriers limiting the access of drugs to their targets. Since characterization of a compound as a transporter substrate or inhibitor bears significant consequences in drug development, there is a great need for reliable tools that enable the rapid analysis of the transport susceptibility of drugs. Here we describe a simple but very efficient high-performance liquid chromatography/mass spectrometry (HPLC/MS) assay for measuring the ABC transporter-dependent vesicular transport of compounds. In addition, we provide evidence that the requirement for sample preparation can be minimized using desorption electrospray ionization (DESI)-MS, paving the way for a direct, high-throughput investigation of drug-transporter interactions.
Institute of Enzymology, Hungarian Academy of Sciences, H-1113 Budapest, Hungary.
Mária
M
Katona
Katalin
K
Kiss
Vilmos
V
Angyal
Nóra
N
Kucsma
Balázs
B
Sarkadi
Zoltán
Z
Takáts
Gergely
G
Szakács
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008727:Methotrexate / Q000493:pharmacokinetics; D009363:Neoplasm Proteins / Q000378:metabolism; D010599:Pharmacokinetics; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D022161:Transport Vesicles / Q000378:metabolism
C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D009363:Neoplasm Proteins; D008727:Methotrexate