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. 2024 Feb 17;35(3):324–332. doi: 10.1021/acs.bioconjchem.3c00519

Figure 3.

Figure 3

Schematic representation of the limited proteolysis protocol. Immunoconjugate, denatured at 68 °C during reduction and alkylation steps, followed three main pathways of unfolding. Pathway A: the only CH2 domain is denatured and fully digested, undigested FabHC, CH3 domain fragments, and intact LC are generated; pathway B: CH2, VH, and CL domains are denatured and fully digested, generating CH3, CH1, and VL single-domain fragments; pathway C: the entire immunoconjugate is denatured and fully digested. Picture created with BioRender.com.