Polypeptide and bacterin for preventing and curing infection of polytype human being papilomavirus
A use and antibody technology, applied in the field of immunology, can solve the problems that HPV has no preventive or therapeutic effect, the vaccine cannot target multiple types of HPV at the same time, and there are few cross-reactions between types
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Embodiment 1
[0064] Preparation of specific peptide antiserum
[0065] (a) Synthesis of HPV L1 short peptide
[0066] In this example, based on the protein sequence of the HPV16L1448-477aa segment, in order to target multiple types of HPV, a G→A mutation was introduced at position 465.
[0067] The designed HPV L1 short peptide is 30 amino acids in length, and its sequence is as follows:
[0068] N-terminal-EVNLKEKFSADLDQFPLARKFLLQAGLKAK-C-terminal (SEQ ID NO: 1).
[0069] Beijing Saibaisheng Gene Technology Co., Ltd. was entrusted to synthesize the HPV L1 short peptide in the Genemed Synthesis Inc. company of the United States by conventional methods.
[0070] (b) Preparation of antiserum
[0071] Dissolve the short peptide synthesized above in PBS, add Furend.s complete adjuvant 1:1, emulsify and immunize Japanese long-eared white rabbits and Balb / c mice with doses of 25ug and 10ug, respectively, three times in total. On the 14th day of the last immunization, the animals were sacrifi...
Embodiment 2
[0073] ELISA Detection of Immune Specificity of Short Peptide Immune Antiserum
[0074] HPV16 virus particles, recombinant HPV16L1VLP, HPV6b L1VLP, HPV16 virus particle mouse antiserum, recombinant HPV16L1VLP and HPV6b L1VLP rabbit and mouse antisera were obtained from the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences; Disease Research Institute. HRP-labeled and fluorescent-labeled secondary antibodies were purchased from Beijing Zhongshan Biotechnology Co., Ltd. (Sigma products). Both Caski cells and Hela cells were purchased from the China Center for Type Culture Collection (CCTCC) of Wuhan University.
[0075] Coat HPV16 virus, recombinant HPV16L1VLP, HPV6b L1VLP, Caski cultured cells containing HPV16 virus, and Hela cultured cell lysates containing HPV18 or their culture fluids on plastic plates, and use rabbit or mouse anti-short peptide antiserum to it react.
[0076] The result is as figure 1 shown. The short peptide immune antiserum of m...
Embodiment 3
[0078] Western blot detection of immunospecificity of short peptide immune antiserum
[0079] Caski cells, Hela cells, and insect cells cultured for 72 hours after transfection with HPV6b and 16L1 recombinant baculovirus were lysed, and the lysate was subjected to SDS polyacrylamide gel electrophoresis. anti.
[0080] The result is as figure 2 shown. In the range of protein molecular weight about 56K, there is a positive reaction band.
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