Abstract
This method describes the conjugation of a synthetic glycopeptide to the N-terminus of a recombinant human interleukin-2 (IL-2) protein fragment. The IL-2 protein fragment is produced as an affinity-tagged fusion protein in Escherichia coli and then cleaved with the highly selective TEV protease to remove the affinity tag and uncover an N-terminal cysteine. The N-terminal cysteine is then used in native chemical ligation reaction to join the IL-2 protein fragment to a glycosylated tripeptide thioester that had been previously synthesized to produce a glycosylated form of IL-2.
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Tolbert, T.J., Wong, CH. (2004). Conjugation of Glycopeptide Thioesters to Expressed Protein Fragments. In: Niemeyer, C.M. (eds) Bioconjugation Protocols. Methods in Molecular Biology™, vol 283. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-813-7:255
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DOI: https://doi.org/10.1385/1-59259-813-7:255
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-58829-098-4
Online ISBN: 978-1-59259-813-7
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