The study of small drug molecules and their behavior in complex biofluids such as serum and plasma is an essential part of pharmaceutical and DMPK research. These small molecules are studied for such characteristics as steady-state concentrations and serum binding equilibrium, and these studies require the isolation of the molecule of interest from the protein laden sample before analysis. The typical isolation process entails precipitating the protein with an organic solvent such as acetonitrile, and removing the precipitate with either centrifugation or filtration. The 96 Well Protein Precipitation Filter Plate offered by ArcticWhite provides a format for rapid, simple, reproducible, and automatable protein precipitation and sample preparation. When using the recommended precipitation conditions of 1.5-3.0:1 (v/v) of solvent:biofluid, the 2mL per well volume allows processing of 15µL-400µL sample volumes per well.
Features of the ArcticWhite Protein Precipitation Filter Plate
- Unique proprietary treatment of Hydrophobic/Oleophobic filters – retains acetonitrile/sample mix above filters even during full mixing – no leakage.
- Pre-filter retains large flocculent particles – prevents any blockage problems.
- Final filter to trap fine protein particles.
- Ideal flow rate due to pore size of filters – proprietary treatment allows higher pore size to be used than other plates.
- Filtrates are easily collected by vacuum, positive pressure or centrifugation.
- Plate format allows for parallel processing and better automation of separation.
- No protein breakthrough.
- No adsorbing of relevant analytes on inert polyethylene filters.
Material and Accessories:
96 Well Collection Plate:
AWLS-219002 (PP, 96 Round Well, 1mL/well)
AWLS-219006 (PP, 96 Square Well, 350µL/well)
AWLS- 219008 (PP, 96 Square Well, 1.0mL/well)
AWLS- 219009 (PP, 96 Square Well, 2.0mL/well)
Protein Precipitation Protocol
- Add 1.5-3.0 volumes of Acetonitrile to the well in the plate.
- Add 1 volume of sample to the well.
- Place on vortex shaker and vortex for 0.5 to 20 mins. Depending on the volume size you may need a cap for the top of the plate.
- a) For Vacuum Processing: Place sample collection plate to bottom of a vacuum manifold and place the filter plate on the top. Gently turn up the vacuum and leave running until all wells have been drawn through.
b) For Centrifugation: Place sample collection plate to bottom of the filter plate, place in centrifuge. Centrifuge at 3500-5000RPM.
The benefit of this plate is that you can add the acetonitrile to the plate and have no leakage until vacuum or centrifugation is applied.