High-efficiency recombinant protein purification using mCherry and YFP nanobody affinity matrices.

ABSTRACT: Mammalian cell lines are important expression systems for large proteins and protein complexes, particularly when the acquisition of post-translational modifications in the protein's native environment is desired. However, low or variable transfection efficiencies are challenges that must be overcome to use such an expression system. Expression of recombinant proteins as a fluorescent protein fusion enables real-time monitoring of protein expression, and also provides an affinity handle for one-step protein purification using a suitable affinity reagent. Here, we describe a panel of anti-GFP and anti-mCherry nanobody affinity matrices and their efficacy for purification of GFP/YFP or mCherry fusion proteins. We define the molecular basis by which they bind their target proteins using X-ray

SUBMITTER: Cong ATQ 

PROVIDER: S-EPMC9413470 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature