Human papillomavirus L1 protein expressed in Escherichia coli self-assembles into virus-like particles that are highly immunogenic.
HPV vaccines based on L1 virus-like particles (VLPs) provided a high degree of protection against HPVs infection. In this study, the codon optimized HPV16 L1 gene were sub-cloned into five procaryotic expression vectors (pET-28a, pET-32a, pGEX-4T-2, pE-sumo and pHSIE), and fused with different protein tags. No recombinant proteins were expressed in pET-28a-L1 and pHSIE-L1, and the proteins expressed by pET-32a-L1 plasmid with TRX-tag were in the form of inclusion body. Only SUMO-tagged and GST-tagged L1 proteins expressed by pE-Sumo-L1 or pGEX-4T-L1 were soluble. The yield of SUMO-L1 protein reached 260mg/L fermentation medium in shake flask. After SUMO tags were eliminated, a 90% purity of L1 proteins was generated by ion-exchange and Ni-NTA affinity chromatography. The purified HPV16 L1 protein self-assembled into virus-like particles (VLPs) and showed a haemagglutination activity. High titers specific and neutralizing antibodies were detected in HPV 16 L1VLPs vaccinated mice. Cytokines such as IFN-γ and IL-2 showed significant higher in VLPs vaccinated mice compared with negative control (p<0.05, p=0.055). Thus, the expression of recombinant HPV16 L1 VLPs in Escherichia coli was feasible, which could potentially be used for a VLP-based HPV vaccine.
College of Basic Medical Science, Zhengzhou University, Zhengzhou 450001, China; Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Yumei
Y
Chen
Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Yunchao
Y
Liu
College of Basic Medical Science, Zhengzhou University, Zhengzhou 450001, China; Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China. Electronic address: zhanggaiping2003@163.com.
Gaiping
G
Zhang
School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China. Electronic address: pingaw@126.com.
Aiping
A
Wang
College of Basic Medical Science, Zhengzhou University, Zhengzhou 450001, China. Electronic address: dongzm@zzu.edu.cn.
Ziming
Z
Dong
School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China.
Yanhua
Y
Qi
College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China.
Jucai
J
Wang
College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China.
Baolei
B
Zhao
School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China.
Ning
N
Li
School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China.
Min
M
Jiang
D000818:Animals; D057134:Antibodies, Neutralizing / Q000096:biosynthesis*; D000914:Antibodies, Viral / Q000096:biosynthesis*; D036022:Capsid Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000276:immunology; D006386:Hemagglutination Tests; D006387:Hemagglutination, Viral; D052162:Human papillomavirus 16 / Q000187:drug effects / Q000254:growth & development / Q000276:immunology; D006801:Humans; D007114:Immunization; D002479:Inclusion Bodies / Q000737:chemistry; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D007376:Interleukin-2 / Q000096:biosynthesis / Q000276:immunology; D051379:Mice; D009856:Oncogene Proteins, Viral / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D030361:Papillomavirus Infections / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D053918:Papillomavirus Vaccines / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics / Q000378:metabolism; D058425:Vaccines, Virus-Like Particle / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D036022:Capsid Proteins; C114580:HPV L1 protein, Human papillomavirus; D007376:Interleukin-2; D009856:Oncogene Proteins, Viral; D053918:Papillomavirus Vaccines; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D058425:Vaccines, Virus-Like Particle; D007371:Interferon-gamma
Bacterial expression; Human papillomavirus; Neutralizing antibody; Splenic lymphocytes; Virus-like particles