Design of Cell-Penetrating Domain Antibodies via a Genetically Encoded β-Lactam Amino Acid.

ABSTRACT: Domain antibodies such as monobodies provide an attractive immunoglobin fold for evolving high-affinity protein binders targeting the intracellular proteins implicated in cell signalling. However, it remains a challenge to endow cell permeability to these small and versatile protein binders. Here, we report a streamlined approach combining orthogonal crosslinking afforded by a genetically encoded β-lactam-lysine (BeLaK) and genetic supercharging to generate cell-penetrating monobodies. When introduced to the N-terminal β-strand of a series of supercharged monobodies, BeLaK enabled efficient inter-strand crosslinking with the neighbouring lysine. Compared to its non-crosslinked counterpart, a BeLaK-crosslinked, +18-charged monobody exhibited enhanced thermostability and greater cellular upt

SUBMITTER: Rabb JD 

PROVIDER: S-EPMC11850181 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature