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 The first involves affinity maturation by complementarity determining region (CDR) walking based on unpublished crystal structure results from the Wilson and Janda laboratories, while the second relies on random recombination, or deoxyribose nucleic acid (DMA) shuffling.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（由申请方提供）：该提案提出了一种用于治疗可卡因成瘾的免疫策略，其中进化出催化抗体，使其能够将可卡因水解为其非精神活性产物ecognine甲酯和苯甲酸。使用之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单链抗体库。第一个涉及基于Wilson和Janda实验室未发表的晶体结构结果的互补决定区（CDR）步移的亲和力成熟，而第二个依赖于随机重组或脱氧核糖核酸（DMA）改组。将使用高通量方法筛选这些GNLM（GNL-突变体）文库，并评估其水解可卡因的能力;将通过比较其动力学参数来确定前两种抗体。将检查选定的GNLM催化剂影响大鼠运动活性的能力，因为运动行为在标准化条件下提供了灵敏且可重现的药物作用测量。