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Updated: May 8, 2026
Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
[Expression and purification of HPV16 E2 protein and preparation of its antiserum]
Yuhui Sun1, Yanjun Zhang, Mingming Liu
1Department of Microbiology, Harbin Medical University, Harbin, China.
Objective:
To express and purify the human papillomavirus type 16 (HPV16) E2 protein in prokaryotic bacteria and prepare the antiserum of HPV16 E2.
Methods:
After amplified by PCR, HPV16 E2 was inserted into pET21b vector. The recombinant pET21b-HPV16E2 vector was transfected into E.coli BL21 (DE3). Expression product was identified after induction. Through purification, denaturation and renaturation, soluble protein was obtained. With the HPV16 E2 protein, we immunized BALB/c mice and examined mouse IFN-γ, CD4(+); T cells, CD8(+); T cells, CD4/CD8 ratio and antiserum titer.
Results:
Restriction digestion and DNA sequencing showed pET21b-HPV16E2 was constructed successfully. Relative molecular mass (Mr;) of HPV16 E2 was 42 000 in SDS-PAGE and the specificity of the protein was confirmed with Western blotting. The antiserum could specifically bind with HPV16 E2 protein. In the immunized BALB/c mice, antiserum titre, CD4(+); T cell count and CD4/CD8 ratio increased, while mouse IFN-γ did not change obviously.
Conclusion:
Soluble HPV16 E2 protein was obtained successfully. The antiserum of high titer against HPV16 E2 was prepared in mice.
