Virology Laboratory, Advanced Centre for Treatment, Research & Education in Cancer (ACTREC) Tata Memorial Centre, Navi, Mumbai.
Indian J Med Res. 2009 Sep;130(3):213-8.
BACKGROUND & OBJECTIVE: Recombinant DNA technology allows expression of the human papillomavirus (HPV) major capsid protein (L1) in heterologous expression systems and the recombinant protein self assembles to virus-like particles (VLP). We took up this study to produce recombinant HPV-16 L1 in yeast, establish the process of recombinant L1 derived VLP preparation and develop an ELISA using VLP as the antigen for serological evaluation of anti HPV-16 L1 antibody status.
Complete HPV-16 L1 was amplified from genomic DNA of an esophageal cancer biopsy, cloned and the protein was expressed in a galactose-inducible Saccharomyces cerevisiae expression system. Self assembled VLP was purified by a two-step density gradient centrifugation process and the VLP preparation used to test its suitability in developing an ELISA.
The recombinant protein was predominantly a ~55 KD species with distinct immunoreactivity and formed VLP as confirmed by electron microscopy. An ELISA using the VLP showed its efficacy in appropriate immunoreactivity to serum/plasma IgG.
INTERPRETATION & CONCLUSION: Recombinant HPV-16 capsid protein derived VLP was produced and the VLP antigen based ELISA can be used to probe serological association of HPV with different clinical conditions. The VLP technology can be improved further and harnessed for future vaccine development efforts in the country.
重组 DNA 技术可使人类乳头瘤病毒（HPV）主要衣壳蛋白（L1）在异源表达系统中表达，重组蛋白自行组装成病毒样颗粒（VLP）。我们开展这项研究，目的是在酵母中产生重组 HPV-16 L1，建立重组 L1 衍生的 VLP 制备工艺，并开发一种 ELISA，使用 VLP 作为抗原，用于血清学评估 HPV-16 L1 抗体状态。
从食管癌活检的基因组 DNA 中扩增完整的 HPV-16 L1，克隆并在半乳糖诱导的酿酒酵母表达系统中表达该蛋白。通过两步密度梯度离心法纯化自组装的 VLP，并将 VLP 制剂用于测试其在开发 ELISA 中的适用性。
重组蛋白主要为~55KD 物种，具有明显的免疫反应性，并通过电子显微镜证实形成了 VLP。使用 VLP 的 ELISA 显示其在适当的免疫反应性方面对血清/血浆 IgG 有效。
已产生重组 HPV-16 衣壳蛋白衍生的 VLP，基于 VLP 抗原的 ELISA 可用于探测 HPV 与不同临床状况的血清学关联。VLP 技术可以进一步改进，并为该国未来的疫苗开发工作提供支持。
