[Production of human papillomavirus type 16 virus-like particles and its immunogenicity].
HPV16 L1 gene was amplified from HPV16 positive vaginal secretion sample by PCR, and inserted into pTO-T7 to obtain the recombinant expression vector pTO-T7-HPV16-L1. Then, the pTO-T7-HPV16-L1 was transformed into E. coil strain ER2566 and the recombinant protein HPV16 L1 was expressed in soluble form. After purification by ammonium sulfate precipitation, ion-exchange chromatography, and hydrophobic interaction chromatography, the recombinant protein HPV16 L1 had a purity of more than 98%. By removing DTT, purified HPV16 L1 proteins self-assembled in vitro into VLPs with the diameter of 50 nm. The vaccination experiments on experimental animals showed the VLPs could elicit high titer of neutralizing antibodies against HPV 16. HPV16 VLPs with high immunogenicity and high purity can be produced easily and effectively from an E. coli expression system in the study, and thus can be used in structure investigation and HPV16 vaccine development.
Wei
Min-Xi
MX
National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen 361005, China. wmx@xmu.edu.cn
Li
Shao-Wei
SW
Huang
Bo
B
Shen
Wen-Tong
WT
Su
Yong-Zai
YZ
Zhang
Chun-Hua
CH
Gu
Ying
Y
Du
Hai-Lian
HL
Zhang
Jun
J
Xia
Ning-Shao
NS
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology / Q000648:ultrastructure; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D006041:Goats; D052162:Human papillomavirus 16 / Q000235:genetics / Q000276:immunology* / Q000648:ultrastructure; D006801:Humans; D008297:Male; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000276:immunology* / Q000821:virology; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014611:Vaccination; D014771:Virion / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; C063317:L1 protein, Human papillomavirus type 16