Persistent infection of high-risk human papillomavirus (HPV) is a leading cause of some cancers, including cervical cancer. However, with over 20 carcinogenic HPV types, it is difficult to design a multivalent vaccine that can offer complete protection. Here, we describe the design and optimization of a HPV51/69/26 triple-type chimeric virus-like particle (VLP) for vaccine development. Using E. coli and a serial N-terminal truncation strategy, we created double- and triple-type chimeric VLPs through loop-swapping at equivalent surface loops. The lead candidate, H69-51BC-26FG, conferred similar particulate properties as that of its parental VLPs and comparable immunogenicity against HPV51, −69 and −26. When produced in a GMP-like facility, these H69-51BC-26FG VLPs were verified to have excellent qualities for the development of a multivalent HPV vaccine. This study showcases an amenable way to create a single VLP using type-specific epitope clustering for the design of a triple-type vaccine.

高危人乳头瘤病毒 (HPV) 的持续感染是导致某些癌症（包括宫颈癌）的主要原因。然而，由于有 20 多种致癌 HPV 类型，很难设计出可以提供全面保护的多价疫苗。在这里，我们描述了用于疫苗开发的 HPV51/69/26 三重型嵌合病毒样颗粒 (VLP) 的设计和优化。使用大肠杆菌和连续的 N 末端截断策略，我们通过等效表面环的环交换创建了双重和三重类型的嵌合 VLP。主要候选者 H69-51BC-26FG 具有与其亲代 VLP 相似的颗粒特性以及与 HPV51、-69 和 -26 相当的免疫原性。当在类似 GMP 的设施中生产时，这些 H69-51BC-26FG VLPs 被证实具有用于开发多价 HPV 疫苗的优良品质。本研究展示了一种使用特定类型表位聚类来创建单个 VLP 的可行方法，用于设计三联疫苗。