Development of a High-Throughput Format for Solid-Phase Extraction of Enantiomers Using an Immunosorbent in 384-Well Plates

摘要

An antibody-based solid-phase extraction method for filtered 384-well plates was developed for a medical drugcandidate having two enantiomeric forms in order to demonstrate the pot6ntial of the use of recombinant antibody fragments as specific and efficient immunosorbents. An immobilization method using a six-histidine tag of the antibody fragment and mild oxidation was applied in order to immobilize antibody fragments in an oriented and ldnetically stable way that ensured high capacity of the antibody support. Phosphate buffer or plasma spiked with enantiomers were used as samples. Selective solidphase extraction was followed by liquid chromatographymass spectromety analysis. Average recoveries for biber and plasma samples ranged from 79 to 122percent and 80 to108percent, respectiVely. Good linearity was observed in the concentration range of 30-3000 ng/mL of the enantiomer.