@inbook{fb7a36ef86f24c5ba9348356e6f1430f,
title = "E6 Tagged Protein Production, Extraction, and Purification from Escherichia coli Lysate",
abstract = "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.",
keywords = "Affinity chromatography, Cervical cancer, E6 protein, Fusion protein, His-tag, Maltose-binding protein, Protein expression and purification, Size exclusion chromatography, Western blot",
author = "Diana Gomes and Passarinha, {Lu{\'i}s A.} and {\^A}ngela Sousa",
note = "Funding Information: info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F00709%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F00709%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT# The authors acknowledge the support from FEDER funds through the POCI-COMPETE 2020–Operational Programme Competitiveness and Internationalisation in Axis I–Strengthening Research, Technological Development and Innovation (Project POCI-01-0145-FEDER-007491), from FCT–Funda{\c c}{\~a}o para a Ci{\^e}ncia e Tecnologia. Associate Laboratory Institute for Health and Bioeconomy–i4HB (project LA/P/0140/2020) which are financed by National Funds from FCT/MCTES. D. Gomes also acknowledges the doctoral fellowship from FCT (ref: 2020.06792.BD). Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2023",
month = apr,
day = "25",
doi = "10.1007/978-1-0716-3147-8_2",
language = "English",
isbn = "978-1-0716-3146-1",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "21--33",
editor = "{\^A}ngela Sousa and Lu{\'i}s Passarinha",
booktitle = "Advanced Methods in Structural Biology",
address = "United States",
}
