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A comprehensive literature search covering the period from 1991 to 2024 was conducted using multiple electronic databases, including PubMed, Web of Science, Scienceing, Google Scholar, Elsevier, CNKI, WanFang, and WeiPu

In addition to the model protein, a Her2-targeting nanobody, we also successfully produced a RNF43-specific nanobody with an N-terminal Gln (N-Gln), an EGFR-targeting nanobody with N-Ala, the Sortase A enzyme with N-Gln, the fluorescent protein TurboGFP with N-Glu, and the sialic acid transferase 15 Pd2,6ST with N-Cys