ANTIBODY CATALYZED CLEAVAGE OF CELL WALL SUBSTRATES

抗体催化细胞壁底物的裂解

• 批准号: 6169347
• 负责人: JEFFREY C GILDERSLEEVE
• 金额: $ 3.24万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-28 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6169347
• 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.

细菌中出现的抗生素耐药性具有严重的医学意义 后果。拟议工作的目标是评估使用 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-丙氨酸-D-紫胶的壁基板。这份提案概述了一个 构建单链抗体噬菌体展示文库的策略 然后选择催化分解酯的特定抗体 D-丙氨酸-D-乳酸链。这些抗体将被检测出它们的 使用标准微量稀释液阻止细胞壁生物合成的能力 肉汤化验。此外，与D-丙氨酸-D-乳酸结合的抗体也将 从噬菌体文库中挑选并在微量稀释液中进行测试 化验。对这两种抗体的比较应有助于 评价催化与结合抑制的优势 生化过程。