Cancer Research Institute, Central South University, Changsha, Hunan, China.
Mol Cell Biochem. 2012 Jun;365(1-2):77-84. doi: 10.1007/s11010-012-1245-7.
Epidermal growth factor-like domain 7 (EGFL7) has been implicated in promoting solid tumor growth and metastasis via stimulating tumor-associated angiogenesis. The advent of antibody display technology (phage, bacteria, and yeast) led to an enormous revival in the use of antibodies as diagnostic and therapeutic tools for fighting cancer. However, problems with protein folding, posttranslational modification, and codon usage still limit the number of improved antibodies that can be obtained. We describe here the isolation of an EGFL7-specific antibody from a mammalian cell-based full-length antibody display library generated from peripheral blood mononuclear cells of patients with hepatocellular carcinoma. Using a novel vector, contained glycosylphosphatidylinositol anchor and restriction enzyme sites NheI and ClaI, antibody libraries are displayed as whole IgG molecules on the cell surface and screened for specific antigen binding by a combination of magnetic beads and measured by cell ELISA. Anti-EGFL7 antibody was successfully isolated from the library. The mammalian cell-based full-length antibody display library is a great potential application for rapid identification and cloning of human mAbs of targeting hepatocellular carcinoma.
表皮生长因子样结构域 7(EGFL7)已被证明可通过刺激肿瘤相关血管生成来促进实体瘤的生长和转移。抗体展示技术(噬菌体、细菌和酵母)的出现极大地促进了抗体作为诊断和治疗癌症工具的应用。然而,蛋白质折叠、翻译后修饰和密码子使用等问题仍然限制了可以获得的改进抗体的数量。我们在这里描述了一种从肝癌患者外周血单个核细胞生成的基于哺乳动物细胞的全长抗体展示文库中分离 EGFL7 特异性抗体的方法。使用一种新型载体,该载体包含糖基磷脂酰肌醇锚和限制酶 NheI 和 ClaI 位点,抗体文库作为完整的 IgG 分子展示在细胞表面,并通过磁珠组合筛选特定的抗原结合,通过细胞 ELISA 进行测量。成功地从文库中分离出抗 EGFL7 抗体。基于哺乳动物细胞的全长抗体展示文库是快速鉴定和克隆针对肝癌的人源单克隆抗体的一种极具潜力的应用。