CATALYTIC ANTIBODIES FOF GAR TRANSFORMYLATION
CATALYTIC ANTIBODIES FOF GAR TRANSFORMYLATION
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
中文摘要
这项提案概述了开发催化抗体的战略。
GAR转化。营养缺乏性互补将被用于
筛查催化抗体,因为筛查是基于催化
活跃性和非约束性。由于GARTfase的底物位于
细胞质,将有必要表达组合文库
(来自用多底物抑制剂免疫的小鼠
β-TGDF)。日本血吸虫功能性抗体的产生
利用硫氧还蛋白还原酶和硫氧还蛋白还原酶的突变体将促进细胞质的形成
利用在细胞质中表达的抗体优化orotate
本科维奇博士的实验室以前描述的是脱羧基。这个
最近体内的紧急情况(功能性抗体表达于
真核细胞质)作为治疗分子展示了
探索功能性抗体生产的重要性
细胞质。GARTfase催化抗体将有益于
表征了GAR转化的酶学性质。这
这些知识将有助于抗肿瘤药物的设计
抑制GARTfase。除了这一重要贡献之外,
该项目将产生催化抗体,酶的性质是琼脂
转化和细胞质抗体的产生,这个项目
会让申请者在组合图书馆有宝贵的经验
构建,基因组操作,催化抗体策略,
酵母菌的酶特性、抗体表达及优化
大肠埃希菌。
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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海外基金