有效抗肿瘤疫苗的开发是癌症预防/免疫治疗领域的重要方向。对于诱导有效的抗肿瘤反应而言，高效的抗原传递至关重要。来自布鲁氏菌属的类酪氨酸合成酶（BLS）是一种具有传递和佐剂特性的十聚体蛋白质，但其在肿瘤疫苗中的应用有限。在这里，我们利用SpyCatcher/SpyTag的Plug-and-Display系统，通过将BLS不对称组装和抗原传递平台相结合，开发了一种抗原传递平台。构建了一个由BLSke和BLSdr组成的不对称组装系统，以等量地组装两个分子。然后，利用SpyCatcher/SpyTag系统将MHC-I限制的卵白素肽（OVA（257-264）SIINFEKL）与BLSke结合，将细胞穿透肽（CPP）KALA与BLSdr结合。KALA修饰增强了OVA肽段进入DC的内部化，促进了DC的成熟和SIINFEKL的交互呈递。此外，在E.G7-OVA肿瘤负载小鼠中，KALA修饰的疫苗免疫治疗抑制了肿瘤生长并增强了CD8+ T细胞反应。在预防模型中，KALA修饰的疫苗接种显示出最显著的保护效应，并显著延长了受挑战小鼠的生存期。总之，不对称组装平台可以等量地组装两种蛋白质或肽段，避免它们的空间或功能干扰。这种不对称组装和Plug-and-Display技术提供了一个快速开发个性化肿瘤疫苗的通用平台。

The development of effective tumor vaccines is an important direction in the field of cancer prevention/immunotherapy. Efficient antigen delivery is essential for inducing effective antitumor responses for tumor vaccines. Lumazine synthase (BLS) from Brucella spp. is a decameric protein with delivery and adjuvant properties, but its application in tumor vaccines is limited. Here, we developed an antigen delivery platform by combining a BLS asymmetric assembly and the Plug-and-Display system of SpyCatcher/SpyTag. An asymmetric assembly system consisting of BLSke and BLSdr was developed to equally assemble two molecules. Then, the MHC-I-restricted ovalbumin peptide (OVA(257-264) SIINFEKL) was conjugated with BLSke, and a cell-penetrating peptide (CPP) KALA was conjugated with BLSdr using the SpyCatcher/SpyTag system. KALA modification enhanced internalization of OVA peptides by DCs as well as promoted the maturation of DCs and the cross-presentation of SIINFEKL. Moreover, the immunotherapy of a KALA-modified vaccine suppressed tumor growth and enhanced CD8+ T cell responses in E.G7-OVA tumor-bearing mice. In the prophylactic model, KALA-modified vaccination showed the most significant protective effect and significantly prolonged the survival period of tumor challenged mice. In conclusion, the asymmetric assembly platform equally assembles two proteins or peptides, avoiding their spatial or functional interference. This asymmetric assembly and Plug-and-Display technology provide a universal platform for rapid development of personalized tumor vaccines.