6.

Simulation of random reduction reaction. cAC10 reduction was simulated with equal rate constants for reduction of heavy-heavy and heavy-light disulfides. (A) The simulated drug loading abundance is shown with bars for the reduction of an average of 2 disulfides to yield an average of 4 antibody cysteines. The percent differences between the experimentally determined drug loading abundance from HIC-HPLC data for DTT and TCEP partial reduction shown in Figure 2 and the simulated values are shown in lines. (B) The simulated isomer distribution is shown in bars. The percent differences between the experimentally determined isomer abundances shown in Figure 5 and the simulated values are shown in lines. In both panels, a positive number indicates that the experimental data is greater than the simulated value, while a negative number indicates that the experimental data is lower than the simulated value.