Purification of the NCOA4 Protein via Liquid Chromatography (2025)

The NCOA4 protein has been shown to play an important role in Ferritinophagy, a form of autophagy, where NCOA4 acts as a cargo receptor and participates in ferroptosis. Recently NCOA4 has attracted increasing attention in cancer treatment with the idea that ferroptosis modulation via NCOA4 could be a potential treatment for some cancers. Ideally NCOA4 could be used to selectively induce ferroptosis in cancer cells effectively targeting exclusively cancer cells for autophagy. The purpose of this research is to develop a method of isolation and purification to obtain a pure NCOA4 protein via liquid chromatography. To achieve this, an Akta Go liquid chromatography instrument can be used to perform either protein affinity, size exclusion, or ion exchange chromatography experiment. In this research project we use primarily protein affinity with a His-Trap column to purify our His-Tag NCOA4 protein. The eluted fractions are then run on an SDSPAGE gel to determine purity and confirm the presence of the protein.