A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University and N.I. Vavilov Institute of General Genetics, Russian Academy of Science, Moscow 119992, Russia.
Virology. 2010 Nov 10;407(1):7-13. doi: 10.1016/j.virol.2010.08.005.
Human epidermal growth factor receptor-2 (HER2/neu) is a target for the humanized monoclonal antibody trastuzumab. Recently, trastuzumab-binding peptides (TBP) of HER2/neu that inhibit proliferation of breast cancer cells were identified. We have now studied conditions of efficient assembly in vivo of Tobacco mosaic virus (TMV)-based particles displaying TBP on its surface. The system is based on an Agrobacterium-mediated co-delivery of binary vectors encoding TMV RNA and coat protein (CP) with TBP in its C-terminal extension into plant leaves. We show how the fusion of amino acid substituted TBP (sTBP) to CP via a flexible peptide linker can improve the manufacturability of recombinant TMV (rTMV). We also reveal that rTMV particles with exposed sTBP retained trastuzumab-binding capacity but lost an anti-HER2/neu immunogenic scaffold function. Mouse antibodies against rTMV did not recognize HER2/neu on surface of human SK-BR-3 cells.
人表皮生长因子受体-2（HER2/neu）是针对人源化单克隆抗体曲妥珠单抗的靶点。最近，鉴定出了抑制乳腺癌细胞增殖的 HER2/neu 的曲妥珠单抗结合肽（TBP）。我们现在已经研究了在体内有效组装展示 TBP 表面的烟草花叶病毒（TMV）基颗粒的条件。该系统基于农杆菌介导的将编码 TMV RNA 和外壳蛋白（CP）的二元载体与 TBP 递送至植物叶片中的共传递。我们展示了如何通过柔性肽接头将氨基酸取代的 TBP（sTBP）融合到 CP 上来提高重组 TMV（rTMV）的可制造性。我们还揭示了具有暴露的 sTBP 的 rTMV 颗粒保留了曲妥珠单抗结合能力，但失去了抗 HER2/neu 免疫原性支架功能。针对 rTMV 的小鼠抗体不能识别人 SK-BR-3 细胞表面的 HER2/neu。
