Generation of TRAIL-receptor 1-specific human monoclonal Ab by a combination of immunospot array assay on a chip and human Ab-producing mice
Generation of TRAIL-receptor 1-specific human monoclonal Ab by a combination of immunospot array assay on a chip and human Ab-producing mice
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DOI：
10.1002/eji.201040551
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发表时间：
2010-12-01
影响因子：
5.4
通讯作者：
Muraguchi, Atsushi
中科院分区：
文献类型：
--
作者：
Jin, Aishun;Ozawa, Tatsuhiko;Muraguchi, Atsushi
单克隆抗体因其特异性强、毒性低等优点，在恶性肿瘤的免疫治疗中具有广阔的应用前景。在单克隆抗体的临床应用中，单克隆抗体的免疫原性是主要关注的问题之一。我们已经开发了使用具有微孔阵列的芯片的芯片上的免疫斑点阵列测定（ISAAC）技术[1]，其能够直接鉴定来自人外周血淋巴细胞的Ag特异性Ab分泌细胞（ASC）和Ab cDNA的快速克隆，从而导致容易、有效和快速地生产人mAb。然而，用Ag免疫志愿者的局限性阻碍了其用于生产针对所需Ag的人mAb的应用。在这项研究中，我们使用ISAAC方法与含有编码整个人免疫球蛋白H链和kL链基因座的人染色体片段的TransChromo（TC）小鼠[2]组合，用于分离对肿瘤坏死因子相关凋亡诱导配体（TRAIL）受体1型特异性的全人mAb，目的是进行癌症免疫治疗。TRAIL受体在多种肿瘤细胞类型以及正常细胞类型中表达。TRAIL通过激活两种功能性受体，即TRAIL-1和TRAIL-2，在多种人类癌细胞系中诱导凋亡。
Monoclonal antibodies have attracted attention clinically as promising therapeutic agents for immunotherapy against various malignancies because of their high specificity and low toxicity. In the clinical application of mAb, immunogenicity of mAb is one of the major concerns. We have developed immunospot array assay on a chip (ISAAC) technology using chips with an array of microwells [1], which enables direct identification of Ag-specific Ab-secreting cells (ASC) from human peripheral blood lymphocytes and rapid cloning of Ab cDNA, leading to easy, efficient and rapid production of human mAb. However, the limitation of immunizing volunteers with Ag hampers its application for the production of human mAb for desired Ag. In this study, we used the ISAAC method in combination with TransChromo (TC) mice that contain human chromosome fragments encoding the entire human immunoglobulin H-chain and kL-chain loci [2] for isolating fully human mAb specific to TNF-related apoptosis-inducing ligand (TRAIL)-receptors type 1 with the object of cancer immunotherapy. TRAIL receptors are expressed in a wide variety of tumor cell types as well as normal cell types. TRAIL induces apoptosis in a wide variety of human cancer cell lines by activating two functional receptors, TRAIL-