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 The first involves affinity maturation by complementarity determining region (CDR) walking based on unpublished crystal structure results from the Wilson and Janda laboratories, while the second relies on random recombination, or deoxyribose nucleic acid (DMA) shuffling.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）：该提案提出了一种治疗可卡因成瘾的免疫治疗策略，其中催化抗体因其将可卡因水解为其非精神活性产物、ecognine甲酯和苯甲酸的能力而进化。使用先前鉴定的可卡因催化剂（称为 GNL）作为起点，将应用两种不同的定向进化技术来构建新型 scFv 库。第一个涉及根据 Wilson 和 Janda 实验室未发表的晶体结构结果通过互补决定区 (CDR) 步行实现亲和力成熟，而第二个则依赖于随机重组或脱氧核糖核酸 (DMA) 改组。这些 GNLM（GNL 突变体）文库将使用高通量方法进行筛选，并评估其水解可卡因的能力；前两种抗体将通过比较它们的动力学参数来确定。将检查选定的 GNLM 催化剂影响大鼠运动活动的能力，因为运动行为提供了标准化条件下药物作用的灵敏且可重复的测量。