Cocaine degradation: Improving antibody efficacy

可卡因降解：提高抗体功效

• 批准号: 7053385
• 负责人: KATHLEEN M MCKENZIE
• 金额: $ 5.04万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7053385
• 关键词: DNA; abzyme; antibody; behavior test; benzoates; biotransformation; cocaine; drug addiction; high performance liquid chromatography; high throughput technology; hydrolysis; laboratory rat; mutant; polymerase chain reaction; postdoctoral investigator; psychomotor function

DESCRIPTION (provided by applicant): This proposal presents an immunotherapeutic strategy for the treatment of cocaine addiction in which catalytic antibodies are evolved for their ability to hydrolyze cocaine to its non-psychoactive products, ecognine methyl ester and benzoic acid. Using previously identified cocaine catalysts, termed GNL, as a starting point, two different directed evolution techniques will be applied for the construction of novel scFv libraries. The first involves affinity maturation by complementarity determining region (CDR) walking based on unpublished crystal structure results from the Wilson and Janda laboratories, while the second relies on random recombination, or deoxyribose nucleic acid (DMA) shuffling. These GNLM (GNL-mutant) libraries will be screened using high throughput methodology and assessed for their ability to hydrolyze cocaine; the top two antibodies will be determined by a comparison their kinetic parameters. Selected GNLM catalysts will be examined for their ability to affect locomotor activity in rats, since motor behavior provides a sensitive and reproducible measure of drug action under standardized conditions.

描述（由申请人提供）：该提案为可卡因成瘾提供了一种免疫治疗策略，在该策略中，催化抗体的进化而进化，因为它们能够将可卡因水解为其非精神活性产物，Ecognine甲基酯和苯甲酸。使用先前鉴定的可卡因催化剂（称为GNL）作为起点，将使用两种不同的定向进化技术来构建新型SCFV库。第一个涉及通过互补性确定区域（CDR）步行而基于未发表的晶体结构而产生的亲和力成熟，而第二个实验室则依赖于随机重组或脱氧核糖核酸（DMA）改组。这些GNLM（GNL突变）文库将使用高吞吐量方法进行筛选，并评估其水解可卡因的能力。前两种抗体将通过比较其动力学参数来确定。由于运动行为在标准化条件下提供了一种敏感且可重复的药物作用量度，因此将检查选定的GNLM催化剂的影响大鼠运动活性的能力。