High-Throughput Assay for Prediction of Plasma Protein Binding

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The 20 min high-throughput assay estimates the binding of drugs to human serum albumin (HSA) and a1-acid glycoprotein (AGP) to predict the plasma protein binding.

The ready-to-go assay kit provides precise predictions of plasma protein binding in a highly controlled and reproducible assay environment.

Off-the-shelf Plasma Protein Prediction Assay Kit

Sovicell has developed an ideal ready-made assay for predicting plasma protein binding – the TRANSIL PPB Binding Kit.

The rapid, high-throughput assay kit for prediction of plasma protein consists of ready-to-use 96-well microtiter plates. Each plate can be used for measuring HSA and AGP binding of up to 12 compounds.

Rapid and economical assay to predict plasma protein binding of drug candidates

Human serum albumin (HSA) and human a1-acid glycoprotein are the most important plasma binding proteins. Under its TRANSIL trademark Sovicell has developed binding assays for both proteins, the TRANSIL HSA Binding Kit and the TRANSIL AGP Binding Kit.

With TRANSIL HSA and AGP beads binding measurements are equivalent to dialysis with these proteins. The TRANSILXL PPB Binding Kit utilises immobilized HSA and AGP in a physiological ratio of 24 to 1 and is designed as a rapid and economical assay to predict plasma protein binding of drug candidates. Differences in relation to plasma binding arise through variations in plasma composition, due to lipids blocking binding sites in native plasma, and on occasion due to binding to other plasma proteins with low abundance.

Estimating the binding of drugs to HSA and AGP with the five-step assay kit

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

The TRANSIL HSA or
AGP Binding bead with proteins
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:

  • Fraction bound to HSA and AGP
  • Fraction bound to plasma

Benefits of the High-Throughput Assay for Prediction of Plasma Protein

  • Requires only 20 minutes total assay time
  • Accurately measures the affinity of drug candidates to HSA and AGP in a physiological mixture to predict it’s free fraction in plasma
  • Ready-to-go 96-well microtiter plate format generating reliable highly reproducible results and robust correlation to equilibrium dialysis method.
  • Fully quality-controlled albumin and a1-acid glycoprotein binding estimates.
  • Rapid compound quantification due to immobilized plasma proteins
  • The Assay 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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