Structural highlights
Publication Abstract from PubMedThe unique characteristics of nanobodies make them ideal candidates for diagnostic and therapeutic applications. Introducing additional disulfide bonds is one of the most effective ways to enhance a nanobody's stability. However, it always negatively affects the nanobody's affinity and expression in host cells. In this study, an anticarbaryl nanobody (NbCBR) was used as a model to investigate the strategy of additional disulfide bond introduction. The crystal structure of the NbCBR-carbaryl complex was determined, based on which molecular dynamics simulations and virtual screening were carried out. Seven disulfide bond-introduced mutants were then constructed. Compared with the wild-type nanobody, all the mutants retained good sensitivity. The results demonstrate that introducing disulfide bonds between two beta-sheets or anchoring the flexible CDR3 to a beta-sheet can significantly enhance the nanobody's stability under thermal, organic solvent, and pH stress conditions. This study provides a theoretical basis for rational engineering of nanobodies to improve their stability, potentially expanding their further applications. An Effective Way to Introduce Disulfide Bonds into Nanobodies: Using an Anticarbaryl Nanobody Structure as a Model.,Liang CX, Liu ML, Guo PY, Fang RY, Hendrickson OD, Zherdev AV, Wang H, Xu ZL, Shen X J Agric Food Chem. 2026 Jul 1;74(25):19995-20004. doi: 10.1021/acs.jafc.6c03602. , Epub 2026 Jun 22. PMID:42328891[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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