Fig. 3.

Functional screening of hBChE variants. In functional screening of hBChE variants, both solid-phase and liquid-phase screening were used. In solid-phase screening, a gel plug that had a positive signal (a yellow color, indicating a positive Ellman reaction) was isolated. To confirm the results from solid-phase screening, virus particles isolated from gel plugs were used to infect cells cultured in 96-well plates to confirm OP resistance. Viruses that were confirmed to be OP-resistant variants were further purified and sequenced to identify the site of mutation. Variants were subcloned, expressed, purified, and studied further in in vitro incubations to characterize their kinetic parameters.