Cooperative folding as a molecular switch in an evolved antibody binder.

ABSTRACT: Designing proteins with tunable activities from easily accessible external cues remains a biotechnological challenge. Here, we set out to create a small antibody-binding domain equipped with a molecular switch inspired by the allosteric response to calcium seen in naturally derived proteins like calmodulin. We have focused on one of the three domains of Protein G that show inherent affinity to antibodies. By combining a semi-rational protein design with directed evolution, we engineered novel variants containing a calcium-binding loop rendering the inherent antibody affinity calcium-dependent. The evolved variants resulted from a designed selection strategy subjecting them to negative and positive selection pressures focused on conditional antibody binding. Hence, these variants contains m

SUBMITTER: Jonsson M 

PROVIDER: S-EPMC11532951 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature