Protein purification with light via a genetically encoded azobenzene side chain.

ABSTRACT: Affinity chromatography is the method of choice for the rapid purification of proteins from cell extracts or culture supernatants. Here, we present the light-responsive Azo-tag, a short peptide comprising p-(phenylazo)-L-phenylalanine (Pap), whose side chain can be switched from its trans-ground state to the metastable cis-configuration by irradiation with mild UV light. Since only trans-Pap shows strong affinity to α-cyclodextrin (α-CD), a protein exhibiting the Azo-tag selectively binds to an α-CD chromatography matrix under daylight or in the dark but elutes quickly under physiological buffer flow when illuminating the column at 355 nm. We demonstrate the light-controlled single-step purification - termed Excitography - of diverse proteins, including enzymes and antibody fragments, with

SUBMITTER: Mayrhofer P 

PROVIDER: S-EPMC11655525 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature