Purification and Characterization of Antibodies in Single-Chain Format against the E6 Oncoprotein of Human Papillomavirus Type 16

In Human Papillomaviruses- (HPV-) associated carcinogenesis, continuous expression of the E6 oncoprotein supports its value as a potential target for the development of diagnostics and therapeutics for HPV cancer. We previously reported that the I7 single-chain antibody fragment (scFv) specific for...

Full description

Saved in:
Bibliographic Details
Published in:BioMed research international 2018-01, Vol.2018 (2018), p.1-9
Main Authors: Amici, C., Affabris, E., Di Bonito, P., Percario, Z., Verachi, F., Accardi, L.
Format: Article
Language:English
Subjects:
Affinity chromatography
Antibodies
Antigens
Binding sites
Cancer
Carcinogenesis
Carcinogens
Cell cycle
Cervical cancer
Cervix
Chains
Cloning
Drug development
E coli
Format
Health aspects
Human papillomavirus
Immunofluorescence
Immunoglobulins
Infections
Nuclear engineering
Nuclear safety
Oncoproteins
Papillomaviridae
Papillomavirus infections
Plasmids
Proteins
Scientific equipment and supplies industry
Tumors
Vaccines
Viral antibodies
Virology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:In Human Papillomaviruses- (HPV-) associated carcinogenesis, continuous expression of the E6 oncoprotein supports its value as a potential target for the development of diagnostics and therapeutics for HPV cancer. We previously reported that the I7 single-chain antibody fragment (scFv) specific for HPV16 E6, expressed as an intrabody by retroviral system, could inhibit significantly the growth of cervical cancer cells in vitro and was even able to reduce tumor development in experimental HPV-related cancer models. Nevertheless, for the development of therapeutic tools to be employed in humans, it is important to achieve maximum safety guarantee, which can be provided by the protein format. In the current study, two anti-16E6 scFvs derived from I7 were expressed in E. coli and purified in soluble form by affinity chromatography. Specificity, sensitivity and stability in physiologic environment of the purified scFvs were demonstrated by binding studies using recombinant 16E6 as an antigen. The scFvs functionality was confirmed by immunofluorescence in cervical cancer cells, where the scFvs were able to recognize the nuclear E6. Furthermore, an antiproliferative activity of the scFvI7nuc delivered in protein format to HPV16-positive cell lines was observed. Our results demonstrate that functional anti-16E6 scFvs can be produced in E. coli, suggesting that such purified antibodies could be used in the diagnosis and treatment of HPV-induced malignancies.
ISSN:2314-6133
2314-6141
DOI:10.1155/2018/6583852