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CATALYTIC ANTIBODIES FOF GAR TRANSFORMYLATION

CATALYTIC ANTIBODIES FOF GAR TRANSFORMYLATION
GAR 转化催化抗体
批准号:
2634584
负责人:
MARC A OSTERMEIER
金额:
$3.02万
依托单位国家:
美国
项目类别:
财政年份:
1998
资助国家:
美国
项目状态:
未结题
起止时间:
1998-01-01 至

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中文摘要
翻译
该提案概述了开发催化抗体的策略, GAR转化。 营养互补将用于 筛选催化抗体,因为筛选是基于催化 活动,而不是约束。 由于GARTfase的底物存在于 在细胞质中,有必要表达组合文库 (来源于用多底物抑制剂免疫的小鼠 β-TGDF)。 功能性抗体的产生 细胞质将使用硫氧还蛋白还原酶中的突变体来促进, 使用在细胞质中表达的乳清酸抗体进行优化 本科维奇博士的实验室之前描述过的脱羧反应 的 最近紧急的胞内抗体(功能性抗体表达在 真核细胞质)作为治疗分子的作用表明, 研究功能性抗体生产的重要性 细胞质 GARTfase催化抗体将有益于 表征GAR转化的酶性质。 这 这些知识将有助于设计抗肿瘤剂, 抑制GARTfase。 除了这些重要贡献之外, 该项目将使催化抗体,酶的性质为GAR 转化,和细胞质抗体的生产,该项目 将使申请人在组合图书馆的宝贵经验, 构建,基因组操作,催化抗体策略, 酶的表征,抗体的表达和优化, E.杆菌
英文摘要
This proposal outlines a strategy for developing catalytic antibodies to GAR transformylation. Auxotrophic complementation will be used to screen for catalytic antibodies since the screening is based on catalytic activity and not binding. Since the substrates for GARTfase reside in the cytoplasm, it will be necessary to express the combinatorial library (derived from a mouse immunized with the multisubstrate inhibitor beta-TGDF) in the cytoplasm. Functional antibody production in the cytoplasm will be facilitated using mutants in thioredoxin reductase and optimized using an antibody expressed in the cytoplasm for orotate decarboxylation previously characterized in Dr. Benkovic's lab. The recent emergency of intrabodies (functional antibodies expressed in the eukaryotic cytoplasm) as therapeutic molecules demonstrates the importance of exploring functional antibody production in the cytoplasm. GARTfase catalytic antibodies will be beneficial for characterizing the enzymatic nature of GAR transformylation. This knowledge will be useful in the design of anti-neoplastic agents to inhibit GARTfase. In addition to the important contributions this project wold make to catalytic antibodies, the enzymatic nature f GAR transformylation, and cytoplasmic antibody production, this project would allow the applicant valuable experience in combinatorial library construction, genomic manipulations, catalytic antibody strategies, enzyme characterization, and antibody expression and optimization in E. coli.
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  • 项目类别:
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  • 财政年份:
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  • 负责人:
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  • 项目类别:
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  • 财政年份:
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