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Fluorescent Probe for Drugs-Human Serum Albumin (HSA) Binding Assay
―Powerful tool for drug discovery, R&D and metabolic toxicity assay―
Investigating whether drugs bind to Human Serum Albumin (HSA) and identifying the drug binding site on HSA are important for pharmacokinetics. BD140 (D4898), developed by Chang et al., is a novel fluorescent probe which binds to HSA drug binding site 2 specifically.1) BD140 is a useful tool to check whether drugs bind to HSA drug binding site 2.
Advantages
- The binding of drugs to HSA drug binding site 2 is detectable.
- Simple and quick assay can be performed.
Application
＜Assay condition＞
Buffer: 1%DMSO in phosphate buffer(pH7.2 - 7.5)
HSA: 20 μM (50 μL/well)
Drugs: each concentration (50 μL/well)
BD140: 20 μM(50 μL/well)
Incubation: 20-25°C for 30 min
Measurement: plate-reader; excitation=365 nm, emission=585 nm
Buffer: 1%DMSO in phosphate buffer(pH7.2 - 7.5)
HSA: 20 μM (50 μL/well)
Drugs: each concentration (50 μL/well)
BD140: 20 μM(50 μL/well)
Incubation: 20-25°C for 30 min
Measurement: plate-reader; excitation=365 nm, emission=585 nm
Binding of BD140 to HSA was not inhibited completely or nearly by warfarin and phenylbutazone, which bind to HSA drug binding site 1, in our result. On the other hand, binding of BD140 to HSA was inhibited sufficiently by ibuprofen, flurbiprofen and triiodobenzoic acid, which bind to drug binding site 2. Thus, BD140 is available for checking whether drugs bind to HSA drug binding site 2.
Reference
- 1) Fluorescent Dye Cocktail for Multiplex Drug-Site Mapping on Human Serum Albumin
