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Analysis of biological function and efficacy of anti EGFR antibodies isolated from screening of human antibody phage display library specific to human renal cell carcinoma

人肾细胞癌特异性抗体噬菌体展示库筛选分离抗EGFR抗体的生物学功能和功效分析

基本信息

批准号：
20591870
负责人：
SHIROKI Ryoichi
金额：
$ 3万
依托单位：
Fujita Health University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2010
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20591870/
关键词：
腫瘍学腎細胞癌抗体治療完全ヒト型抗体腎癌特異抗原

项目摘要

Using the ICOS method, AIMS5 library was screened with human RCC cell lines, Caki-1(2 cases), CCF-RC1(2 cases) and ACHN(1 case). RCC-specific antibodies were chosen by immunostaining using clinical samples. We chose two different antibodies(059-152 and 062-130) that exhibited cancer cell-specific staining without staining normal cells on clinical RCC sample. These antibodies also reacted to the cell surface antigen of the selected human RCC cells. Antigens these two antibodies reacted were isolated from immunoprecipitation with CCF-RC1.Consequently, the recognized antigen by these two antibodies was proved to be epidermal growth factor receptor(EGFR). Isolated two different anti-EGFR antibodies were assessed for their functional activity on cell proliferation using Caki-1.At the low concentration(0.1μg/ml), 059-152 and 062-130 exhibited about 70% cell proliferation. At the high concentration(10μg/ml), our two anti-EGFR antibodies(059-152, 062-130) elicited up to 40 to 60% cell prolifer … More ation. In ADCC assay, 059-152 had about 35% ADCC assay. 062-130 had about 40% ADCC assay. The effects of the Ab on the phosphorylation reaction were examined. 059-152 did not inhibit phosphorylation in any cell lines. 062-130 and Cetuximab inhibited phosphorylation using CCF-RC1 and ACHN.To determine if the effects of these antibodies could be translated to inhibition of tumor growth in vivo, the antibody was given to mice transplanted with human RCC cells. Treatment with both antibodies developed significant inhibition on human RCC tumor growth compared with control group. The degree of growth inhibition, however, were dependent on the antibody, administration schedule and doses. Although, synergistic growth inhibitions were observed in combination with chemotherapeutic agents, reduction of host mice body weights reduction were also noted, which meant the necessity of investigation of adverse effects in combination therapy.We expected these antibodies were candidate therapeutic antibodies. We showed, our anti-EGFR antibodies, especially 062-130 had sufficient effect. Because our antibodies were originated from human phage display system, possibility of cross reaction over animals were thought to be limited. We expect that this antibody will become a therapeutic antibody for RCC in clinical use. Less

采用ICOS方法对AIMS5文库与人RCC细胞系Caki-1（2例）、CCF-RC1（2例）和ACHN（1例）进行筛选。临床标本免疫染色选择rcc特异性抗体。我们选择了两种不同的抗体（059-152和062-130），它们在临床RCC样本中表现出癌细胞特异性染色，而不染色正常细胞。这些抗体也与选定的人肾细胞癌细胞表面抗原发生反应。这两种抗体反应的抗原是从CCF-RC1免疫沉淀中分离出来的。结果表明，这两种抗体的识别抗原为表皮生长因子受体（EGFR）。分离的两种不同的抗egfr抗体使用Caki-1评估其对细胞增殖的功能活性。在低浓度（0.1μg/ml）下，059-152和062-130的细胞增殖率约为70%。在高浓度（10μg/ml）下，我们的两种抗egfr抗体（059-152、062-130）可诱导高达40% ~ 60%的细胞增殖率。ADCC检测中，059-152的ADCC检测率约为35%。062-130 ADCC含量约为40%。考察了Ab对磷酸化反应的影响。059-152不抑制任何细胞系的磷酸化。062-130和西妥昔单抗通过CCF-RC1和ACHN抑制磷酸化。为了确定这些抗体的作用是否可以转化为体内肿瘤生长的抑制，将抗体给予移植了人RCC细胞的小鼠。与对照组相比，两种抗体治疗对人RCC肿瘤生长均有显著抑制作用。然而，生长抑制的程度取决于抗体、给药计划和剂量。虽然与化疗药物联合使用观察到协同生长抑制，但也注意到减轻宿主小鼠体重，这意味着有必要研究联合治疗的不良反应。我们预期这些抗体是候选治疗抗体。我们发现，我们的抗egfr抗体，特别是062-130有足够的效果。由于我们的抗体来源于人噬菌体展示系统，因此在动物身上交叉反应的可能性被认为是有限的。我们期望该抗体能成为临床应用的RCC治疗性抗体。少

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Methods for comprehensive identification of membrane proteins recognized by a large number of monoclonal antibodies

DOI：
10.1016/j.jim.2009.09.003
发表时间：
2009-12-31
期刊：
JOURNAL OF IMMUNOLOGICAL METHODS
影响因子：
2.2
作者：
Kurosawa, Gene;Sumitomo, Mariko;Kurosawa, Yoshikazu
通讯作者：
Kurosawa, Yoshikazu

Comprehensive screening for antigens overexpressed on carcinoma via isolation of human mAbs that may be therapeutic

通过分离可能具有治疗作用的人单克隆抗体来全面筛查癌症上过度表达的抗原

DOI：
发表时间：
2008
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Kurosawa G;Akahori Y(24人中2番目);Shiroki R(24人中20番目);Hoshinaga K(24人中21番目);et al
通讯作者：
et al

進行腎癌に対する分子標的薬治療の有効性と有害事象の検討

分子靶向药物治疗晚期肾癌的有效性和不良事件检验