Design and Synthesis of Monobody Variants with Low Immunogenicity.
Naoya IwamotoYukino SatoAsako ManabeShinsuke InukiHiroaki OhnoMotohiro NonakaShinya OishiPublished in: ACS medicinal chemistry letters (2023)
Mirror-image proteins (d-proteins) are promising scaffolds for drug discovery because of their high proteolytic stability and low immunogenic properties. Facile and reproducible processes for the preparation of functional d-proteins are required for their application in therapeutic biologics. In this study, we designed and synthesized a novel monobody variant with two cysteine substitutions that facilitate the synthetic process via sequential native chemical ligations and improve protein stability by disulfide bond formation. The synthetic anti-GFP monobody in this model study exhibited good binding affinity to the target enhanced green fluorescent protein. In vivo administration of the synthetic anti-GFP monobody (l-monobody) to mice induced antidrug antibody (ADA) production, whereas no ADA production was observed following immunization with the mirror-image anti-GFP monobody (d-monobody). These results suggest that the synthetic d-monobody is a non-antibody protein scaffold with low immunogenic properties.
Keyphrases
- drug discovery
- protein protein
- binding protein
- deep learning
- quantum dots
- small molecule
- amino acid
- dna methylation
- machine learning
- living cells
- metabolic syndrome
- mass spectrometry
- adipose tissue
- copy number
- endothelial cells
- molecularly imprinted
- dna binding
- highly efficient
- fluorescent probe
- visible light
- wild type