Binding on Silica Beads Immobilized Protein Fraction

Binding on Silica Beads Immobilized Protein Fraction [Pg.485]

UV-plate reader and/or LC/MS-MS determine the concentrations of the unbound fraction at varying HSA concentration. [Pg.485]

This method is based on the affinity of the new chemical entity to a membrane, consisting of human serum albumin immobilized on Transil beads. The silica beads are coated with egg yolk phosphatidylcholine and are commercially available under the trademark Transil from Nimbus (Nimbus Leipzig Germany). [Pg.485]

The plates are delivered in frozen state filled with a total volume of 270 xL of buffer for control well (column 1 and 7) and 270 pL of buffer/varying amounts of Transil for sample well. The plates are slowly defrosted immediately before measurement. [Pg.485]

To each well (control and sample) 30 pL of compound solution has to be added. If UV spectroscopy method is used for quantification the wells H7-H12 have to use as reference sample to account for the absorbance of the UV-plate (not necessary for LC/MS-MS). [Pg.485]

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