Purification and immunogenicity study of human papillomavirus 58 virus-like particles expressed in Pichia pastoris
Purification and immunogenicity study of human papillomavirus 58 virus-like particles expressed in Pichia pastoris
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DOI：
10.1016/j.pep.2011.07.009
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发表时间：
2011-12-01
影响因子：
1.6
通讯作者：
Lou, Jueren
中科院分区：
文献类型：
--
作者：
Jiang, Zijun;Tong, Guangjie;Lou, Jueren
目前市场上有两种人乳头瘤病毒（HPV）预防性疫苗：Gardasil和Cervarix。这两种疫苗针对肿瘤高危亚型HPV 16和HPV 18。然而，它们不包括其他高危亚型，如HPV 58。我国流行病学研究表明，HPV 58是一种流行的高危亚型，仅次于HPV 16和HPV 18。因此，对于中国的宫颈癌预防来说，开发针对HPV 58的疫苗是必要的。在本研究中，HPV 58病毒样颗粒（VLP）在毕赤酵母中表达，并随后通过预处理和三步纯化过程，包括强阳离子交换层析，尺寸排阻层析和羟基磷灰石层析纯化。经十二烷基硫酸钠聚丙烯酰胺凝胶电泳、电子显微镜、动态激光散射和超离心鉴定，证实了高度纯化的HPV 58 VLP。纯化的VLP用于免疫小鼠以测试其诱导体液免疫的能力。对免疫小鼠的血清进行酶联免疫吸附测定，观察到显着高的抗HPV 58 VLP抗体滴度。免疫原性研究表明，纯化的HPV 58 VLP是HPV疫苗候选物。(C)2011 Elsevier Inc. All rights reserved.
Two human papillomavirus (HPV) prophylactic vaccines are currently available in the market: Gardasil and Cervarix. These two vaccines work against tumor high-risk subtypes HPV 16 and HPV 18. However, they do not include other high-risk subtypes such as HPV 58. Epidemiological research in China shows that HPV 58 is a prevalent high-risk subtype, second only to HPV 16 and HPV 18. Thus, for cervical cancer prevention in China, developing a vaccine against HPV 58 is necessary. In this study, HPV 58 virus-like particles (VLPs) were expressed in the Pichia pastoris, and subsequently purified through pretreatment and a three-step purification process consisting of strong cation exchange chromatography, size-exclusion chromatography, and hydroxyapatite chromatography. The highly purified HPV 58 VLPs were confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, electron microscopy, dynamic laser scattering, and ultracentrifugation. The purified VLPs were used to immunize mice to test their ability to induce humoral immunity. Enzyme-linked immunosorbent assays were performed on the sera of the immunized mice and significantly high anti-HPV 58 VLP antibody titers were observed. The immunogenicity study demonstrates that the purified HPV 58 VLPs are HPV vaccine candidates. (C) 2011 Elsevier Inc. All rights reserved.