Cell-free production of scFv fusion proteins: an efficient approach for personalized lymphoma vaccines
Cell-free production of scFv fusion proteins: an efficient approach for personalized lymphoma vaccines
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DOI：
10.1182/blood-2006-07-030593
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发表时间：
2007-04-15
期刊：
影响因子：
20.3
通讯作者：
Levy, Ronald
中科院分区：
文献类型：
--
作者：
Kanter, Gregory;Yang, Junhao;Levy, Ronald
每种b细胞淋巴瘤表面独特的免疫球蛋白（Ig）独特型代表了一种理想的肿瘤特异性抗原，可作为治疗性疫苗使用。我们已经使用基于大肠杆菌的无细胞蛋白表达系统，在从b细胞肿瘤中克隆Ig基因的数小时内生产出疫苗。我们证明了一种由独特型单链Fv抗体片段（scFv）组成的融合蛋白与细胞因子（GM-CSF）或免疫刺激肽连接是一种有效的淋巴瘤疫苗。这些疫苗引发了针对肿瘤表面天然Ig蛋白的体液免疫反应，并保护小鼠免受肿瘤攻击，其效力与在哺乳动物细胞中产生的常规Ig并化学偶联于钥匙孔帽螺血青蛋白相当。无细胞大肠杆菌系统为快速生成个体化疫苗提供了平台，从而使临床应用更加有效。
The unique immunoglobulin (Ig) idiotype on the surface of each B-cell lymphoma represents an ideal tumor-specific antigen for use as a therapeutic vaccine. We have used an Escherichia coli-based, cell-free protein-expression system to produce a vaccine within hours of cloning the Ig genes from a B-cell tumor. We demonstrated that a fusion protein consisting of an idiotypic single chain Fv antibody fragment (scFv) linked to a cytokine (GM-CSF) or to an immunostimulatory peptide was an effective lymphoma vaccine. These vaccines elicited humoral immune responses against the native Ig protein displayed on the surface of a tumor and protected mice against tumor challenge with efficacy equal to that of the conventional Ig produced in a mammalian cell and chemically coupled to key-hole limpet hemocyanin. The cell-free E coli system offers a platform for rapidly generating individualized vaccines, thereby allowing much more efficient application in the clinic.