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

使用ICOS方法，用人RCC细胞系，CAKI-1（2例），CCF-RC1（2例）和ACHN（1例）筛选AIMS5库。通过使用临床样品进行免疫染色来选择RCC特异性抗体。我们选择两种不同的抗体（059-152和062-130），它们暴露了癌细胞特异性染色，而无需在临床RCC样品上染色正常细胞。这些抗体也对所选人RCC细胞的细胞表面抗原有反应。抗原通过CCF-RC1免疫沉淀分离出来的这两种抗体。事实证明，这两种抗体被证明是表皮生长因子受体（EGFR）。使用CAKI-1评估了分离的两种不同的抗EGFR抗体在细胞增殖上的功能活性。在低浓度（0.1μg/ml），059-152和062-130表现出约70％的细胞增殖。在高浓度（10μg/mL）下，我们的两种抗EGFR抗体（059-152，062-130）最多引起了40％至60％的细胞增殖……更多。在ADCC分析中，059-152的ADCC分析约为35％。 062-130的ADCC分析约为40％。检查了AB对磷酸化反应的影响。 059-152均未抑制任何细胞系中的磷酸化。 062-130和西妥昔单抗使用CCF-RC1和ACHN抑制磷酸化。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，对用人RCC细胞移植的小鼠给出了抗体。与对照组相比，两种抗体的治疗对人RCC肿瘤生长均显着抑制。然而，生长抑制的程度取决于抗体，给药时间表，尽管观察到协同生长抑制与化学治疗剂的结合，但也发现了减少宿主体重的减少，这意味着对组合治疗中的不良反应进行了必要的研究。这些抗体尤其是候选抗体抗体，尤其是候选抗体抗体。有足够的效果。由于我们的抗体最初来自人类噬菌体显示系统，因此认为对动物进行交叉反应的可能性受到限制。我们预计该抗体将成为一种用于临床使用的RCC的治疗抗体。较少的

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

Analysis of Biological Function of antiEGFR Antibodies Isolated from Screening of Human Antibody Library Specific to Human RCC

人肾细胞癌特异性抗体库筛选分离抗EGFR抗体的生物学功能分析