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. 2023 Feb 21;9(3):476–487. doi: 10.1021/acscentsci.2c01437

Figure 2.

Figure 2

Chemical construction of a full antibody to generate FcCD20-(FabHER2)2SynAb 2. A Bis-PD method for the construction of a full antibody FcCD20-(FabHER2)2 SynAb 2. FcCD205 is sequentially reduced and rebridged with bis-PD(Tz)29. The resulting FcCD20-(Tz)210 is then reacted with FabHER2-BCN 11 to generate FcCD20-(FabHER2)2 SynAb 2 after SEC purification. B Bis-PD(Tz)29 used for SynAb synthesis. C Br2PD-BCN 4 used for SynAb synthesis. D SDS-PAGE analysis of FcCD20-(FabHER2)2 SynAb 2 formation. Lane 1: Ladder. Lane 2: FabHER2-BCN 11. Lane 3: FcCD20-(Tz)210. Lane 4: crude FcCD20-(FabHER2)2 SynAb 2. E UV trace of SEC purification of FcCD20-(FabHER2)2 SynAb 2. F SDS-PAGE analysis of SEC purification of FcCD20-(FabHER2)2 SynAb 2. Lane 1: Ladder. Lane 2–5: Large species. Lane 6–9: FcCD20-(FabHER2)2 SynAb 2. Lane 10–12: FcCD20-(FabHER2)-Tz 12. G LC-MS analysis of FcCD20-(Tz)210. Expected mass: 51246 Da. Observed mass: 51250 Da. H LC-MS analysis of FabHER2-BCN 11. Expected mass: 48141 Da. Observed mass: 48141 Da. I LC-MS analysis of FcCD20-(FabHER2)-Tz 12. Expected mass: 99359 Da. Observed mass: 99367 Da. J LC-MS analysis of FcCD20-(FabHER2)2 SynAb 2. Expected mass: 147472 Da. Observed mass: 147488 Da.