Figure 8.

Antibody reconstitution in 96-well format using a surplus of Fab-Int^N to demonstrate one-pot purification. A subset of the whole 96-well plate is illustrated. A1: Single anti-CD40 Fab Int^N (61.5 kDa). B1: single huFc Int^C (63.5 kDa). A2 to A9: Antibody fragments anti-CD40 Int^N and huFc Int^C were mixed in a ratio 3:1 and PTS was activated by 0.5 mM TCEP. Reconstituted antibodies were separated from non-reconstituted antibodies via Ni²⁺ beads and further treated with DHAA for reoxidation. Reconstituted anti-CD40 antibodies are indicated at 145.7 kDa under non-reduced conditions. A10-A11: Not treated with Ni²⁺ beads after reconstitution reaction, resulting in reconstituted anti-CD40 antibody as well as antibody fragments anti-CD40 Int^N and huFc Int^C. A12 and B12: Antibody fragments anti-CD40 Int^N and huFc Int^C after TCEP treatment without the addition of Ni²⁺ beads and DHAA (control). Gel band at 1 kDa (green): Lower Marker (LM). Blue band below 16 kDa: System Peak. The whole gel panel of the 96-well plate analysis is shown in Figure S12.