ANTIBODY CATALYZED CLEAVAGE OF CELL WALL SUBSTRATES

抗体催化细胞壁底物的裂解

• 批准号: 6372912
• 负责人: JEFFREY C GILDERSLEEVE
• 金额: $ 4.02万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-28 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6372912
• 关键词: abzyme; bactericidal immunity; biotechnology; cell wall; chemical cleavage; chemical synthesis; dipeptides; drug resistance; enzyme linked immunosorbent assay; enzyme substrate; esterase; microorganism growth; protein engineering; vancomycin

The emergence of antibiotic resistance in bacteria has severe medical consequences. The goal of the proposed work is to evaluate the use of catalytic antibodies for blocking cell wall biosynthesis in vancomycin resistant bacteria. These resistant bacteria synthesize a modified cell wall substrate with a terminal D-ala-D-lac. This proposal outlines a strategy to construct a phage displayed library of scFv antibodies and then select specific antibodies that catalyze the cleavage the ester linkage of D-ala-D-lac. These antibodies will be tested for their ability to block cell wall biosynthesis using a standard microdilution broth assay. In addition, antibodies which bind D-ala-D-lac will also be selected from the phage library and tested in the microdilution broth assay. Comparison of the two types of antibodies should be useful in evaluating the advantages of catalysis versus binding for inhibiting biochemical processes.

细菌抗生素耐药性的出现， 后果 拟议工作的目标是评估 阻断万古霉素细胞壁生物合成的催化抗体 耐药细菌 这些耐药细菌合成了一种改良细胞 具有末端D-ala-D-lac的壁基板。 该提案概述了 噬菌体展示单链抗体库构建策略 然后选择催化酯裂解的特异性抗体， D-ala-D-lac的连接。 这些抗体将被测试其 使用标准微量稀释阻断细胞壁生物合成的能力 肉汤测定 此外，结合D-ala-D-lac的抗体也将 从噬菌体库中选择并在微量稀释肉汤中测试 比色法 两种类型抗体的比较应该是有用的， 评估催化与结合抑制的优势 生化过程

项目成果

Development of a high-throughput screen for protein catalysts: application to the directed evolution of antibody aldolases.

• DOI: 10.1002/anie.200352117
• 发表时间: 2003-12
• 期刊: Angewandte Chemie
• 作者: J. Gildersleeve;A. Varvak;S. Atwell;Doug Evans;P. Schultz

Development of a genetic selection for catalytic antibodies.

开发催化抗体的遗传选择。

• DOI: 10.1016/s0960-894x(02)00198-1
• 发表时间: 2002
• 期刊: Bioorganic & medicinal chemistry letters
• 影响因子: 2.7
• 作者: Gildersleeve,Jeff;Janes,Jeff;Ulrich,Helle;Yang,Priscilla;Barbas,Carlos;Schultz,PeterG
• 通讯作者: Schultz,PeterG

A new strategy for the synthesis of glycoproteins.

糖蛋白合成的新策略。

• DOI: 10.1126/science.1089509
• 发表时间: 2004
• 期刊: Science (New York, N.Y.)
• 作者: Zhang,Zhiwen;Gildersleeve,Jeff;Yang,Yu-Ying;Xu,Ran;Loo,JosephA;Uryu,Sean;Wong,Chi-Huey;Schultz,PeterG
• 通讯作者: Schultz,PeterG