Production of human papillomavirus type 6 virus-like particles and detection of its neutralizing antibodies
10.3760/cma.j.issn.0254-5101.2009.01.017
- VernacularTitle:人乳头状瘤病毒6型类病毒颗粒的制备及其中和抗体的检测
- Author:
Huirong PAN
;
Shaowei LI
;
Bo LIU
;
Ziheng ZHU
;
Huanhuan CAO
;
Ying GU
;
Tong CHENG
;
Yingbin WANG
;
Jun ZHANG
;
Ningshao XIA
- Publication Type:Journal Article
- Keywords:
Human papillomavirus type 6;
Escherichia coli expression system;
Purification;
Virus-like particle;
Immunogenicity
- From:Chinese Journal of Microbiology and Immunology
2009;29(1):71-76
- CountryChina
- Language:Chinese
-
Abstract:
Objective To produce human papillomavirus type 6(HPV-6)virus-like particles with Escherichia coli expression system and study its immunogenicity.Methods HPV-6 L1 gene was inserted into pmkaryotic expression vector pTO-T7 and then expressed in Escherichia coli ER2566.The HPV-6 L1 protein was purified by ammonium sulfate precipitation,ion-exchange chromatography,and hydrophobic interaction chromatography.Then the purified HPV-6 L1 self-assembled into virus-like particle after removing 1,4dithiothreitol(DTr).The morphology of the virus-like particles was investigated with dynamic light scatter and transmission electron microscopy,and the immunogenicity was determined with in vitro pseudownons neutralization as8ay.Results HPV-6 L1 was expressed in soluble form in Escherichia coli.Following the removal of DTT,purified HPV-6 L1 protein could assemble into virus-like particles as 25 am in the radius.And the animal immunization test showed HPV-6 virus-like particles can elite hish titer neutralizing antibodies.Conclusion The bacterially expressed HPV-6 L1 VLP is highly immunogenieity and easy to produce.And it can be good candidate of HPV-6 vaccine.
