E6 Tagged Protein Production, Extraction, and Purification from Escherichia coli Lysate

Cervical cancer has been extensively associated with human papillomavirus (HPV) due to the expression of oncoproteins such as E6. This protein can interfere with p53 tumor suppressor activity, blocking apoptosis of abnormal cells. The functional inhibition of E6 protein is a promising therapeutic strategy for HPV-induced cancers. Conducting biointeraction and characterization studies between E6 protein and potential anti-HPV drugs is necessary to obtain large quantities of high-purity and soluble E6 protein. The recombinant production of E6 protein is particularly challenging because it tends to aggregate. One way to circumvent this problem is to use a dual MBP-His₆ tag that can facilitate the expression, proper folding, and solubility of the E6 protein. This chapter outlines effective methods for expressing and obtaining E6 protein with a dual affinity tag by combining different chromatographic methods.