High-Throughput Assay Kit for Albumin (HSA) Binding

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The 20 min high-throughput assay kit estimates the binding of drugs to human serum albumin (HSA) and predicts the plasma protein binding.

The ready-to-go assay kit measures the affinity constant (KD) of
drugs to albumin and hence allows the calculation of albumin binding even under disease and physiological states that alter the albumin content of human serum. In combination with the
TRANSIL AGP Binding Kit it is possible to obtain a precise prediction of plasma protein binding
in a highly controlled and reproducible assay environment.

Off-the-shelf High-Throughput Assay Kit for Albumin (HSA) Binding

Sovicell has developed a dedicated ready-made assay for measuring the affinity of drug candidates to human serum albumin (HSA) to predict it’s free fraction in plasma – the TRANSIL HSA Binding Kit.

The 20 minute, high-throughput HSA assay kit consists of ready-to-go 96-well microtiter plates. Each plate can be used for measuring HSA binding of up to 12 compounds. When used together with the TRANSIL AGP Binding Kit it enables obtaining an accurate prediction of plasma protein binding in a highly controlled and reproducible assay environment.

The TRANSIL HSA Binding Kit

Simulate a broad range of disease conditions

Human serum albumin (HSA) and human a1-acid glycoprotein are the most important plasma binding proteins. TRANSIL binding assays are available for both proteins. The TRANSIL HSA Binding Kit employs immobilized HSA with a random orientation. This ensures that all binding sites are available and that the assay reproduces exactly the binding of drugs to albumin.

Furthermore, the High-throughput HSA Binding Assay Kit can be used as a rapid screen to predict plasma protein binding, and in conjunction with the TRANSIL AGP Binding assay also to measure plasma protein binding and to simulate various disease conditions. Differences in relation to plasma binding arise through variations in plasma composition, due to lipids blocking binding sites in native plasma, and occasionally due to binding to other plasma proteins with low abundance.

Estimating the binding of drugs to Albumin (HSA) with the five-step assay kit

The TRANSIL HSA Binding Assay kit estimates the binding of drugs in only 5 steps:

HSA Binding
The TRANSIL HSA Binding bead with albumin immobilized in random oriention to expose all binding sites.
  1. addition of drug candidate
  2. mixing and incubating for 12 minutes
  3. removal of beads by centrifugation
  4. sampling of supernatant
  5. quantification of the drug candidate

Capabilities

Detection systems:

  • LC/MS/MS
  • Scintillation counting
  • Others

Parameters estimated and predicted:

  • Affinity constant (KD) of drugs to albumin
  • Fraction bound to albumin
  • Fraction to plasma

Features & Benefits of the High-Throughput Albumin (HSA) Binding Assay

  • Fast, requires only 20 minutes total assay time
  • Accurately measures the affinity of drug candidates to human serum albumin (HSA) to predict it’s free fraction in plasma
  • Reliable with highly reproducible results and robust correlation to equilibrium dialysis method.
  • Fully quality-controlled albumin binding estimates.
  • Rapid compound quantification due to immobilized plasma proteins
  • Kit includes a spreadsheet for calculation of final results and traffic light system for data quality rating

For more information please contact Sovicell today.

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