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Figure 1 | BMC Biotechnology

Figure 1

From: Efficient, chemoselective synthesis of immunomicelles using single-domain antibodies with a C-terminal thioester

Figure 1

Flow chart showing the approach explored in this work. Introduction of a pelB leader sequence targets the sdAb-intein-CBD fusion protein to the periplasm, where the presence of disulfide isomerases and an oxidizing environment ensure proper folding and disulfide bond formation of the antibody domain. The chitin binding domain (CBD) allows single-step purification of the fusion protein from the periplasm, where after on-column treatment with MESNA results in intein-mediated cleavage and the formation of single-domain antibody with a reactive C-terminal thioester, which can then be directly applied in subsequent native chemical ligation reactions.

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