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. 2024 Oct 18;33(11):e5176. doi: 10.1002/pro.5176

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© 2024 The Author(s). Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.

This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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(a) AlphaFold2 predicted VHH1‐NKp30 complex. (b) Visualization of VHH1 paratope residues (within 4.5 Å of NKp30). (c) Depiction of the non‐canonical disulfide bond that covalently links CDR3 to Cys55. (d) 3D‐alignment of AlphaFold2 predicted binding modes of parental VHH1‐v1.0 and humanized VHH1‐v1.9. (e) 3D‐alignment of parental VHH1‐v1.0 and fully framework‐humanized VHH1‐v1.10 indicates a possible steric clash of humanized Hallmark residue Trp52 with NKp30. Key residues that were subjected to humanization are indicated in the same color coding as in Figure 1.