Fig. 4
From: Optimization of a nicotine degrading enzyme for potential use in treatment of nicotine addiction
Effects of PEGylation of NicA2. (a) SDS-PAGE gel showing mobility of NicA2 (lane 2), conjugated NicA2 using a 1:7 (lane 3) or 1:20 (lane 4) molar excess of a 10 kDa linear or 20 kDa linear (lanes 5 and 6) NHS-PEG reagent. The mobility of unconjugated NicA2 (52 kDa) is indicated by an arrow. Lane 1 contains MW markers. (b) Increased circulating half-life of PEGylated NicA2. Wt NicA2 or the 3 different PEGylated preparations were dosed i.v. (5 mg/kg) in rats, and concentrations in serum samples retrieved at various time-points (measured by ELISA) plotted as a function of time. (c) Identical amounts of purified wt NicA2 (circles) or NicA2-PEG2 (squares) were added to rat serum containing 50 ng/mL S-(−)-nicotine. Samples were withdrawn, and activity quenched by MeOH addition at the time points indicated. Nicotine concentration was measured by GC and plotted as a function of time