Neutralizing Human Antibodies to Botulinum Toxin A

中和 A 型肉毒杆菌毒素的人类抗体

• 批准号: 6904434
• 负责人: LISA M EUBANKS
• 金额: $ 4.99万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-16 至 2007-06-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6904434
• 关键词: antigen antibody reaction; bioterrorism /chemical warfare; botulinum toxins; botulism; chemoprevention; cytoprotection; cytotoxicity; disease /disorder prevention /control; drug design /synthesis /production; immunologic substance development /preparation; microorganism disease chemotherapy; neutralizing antibody; phage display; postdoctoral investigator; protein binding; protein protein interaction; tissue /cell culture

DESCRIPTION (provided by applicant): Botulinum neurotoxin type A (BoNT A) is one of the most potent toxins to humans and has become an imminent threat in biological warfare. Administered as either a pre- or post-exposure prophylaxis, high affinity neutralizing antibodies can potentially provide protection against the disease botulism. Human single-chain variable fragment antibodies (scFvs) that recognize BoNT A have previously been identified, including one antibody that can almost completely neutralize BoNT A. While extremely promising, it would be beneficial to enhance the affinity of these antibodies towards BoNT A. This will be achieved by subjecting the scFVs to affinity maturation/molecular evolution methods in combination with phage-display. The most promising antibodies will then be converted to a more stable format for further characterization. The ability of the antibodies to protect against the cytotoxic effects of BoNT A will be determined with an in vitro cell-based assay. Additionally, binding affinity and specificity of these antibodies will be investigated using BIAcore. The association rate constant, dissociation rate constant, and equilibrium dissociation constant for antibody-BoNT A binding will be obtained for the evolved antibodies.

描述（由申请人提供）：A型肉毒杆菌神经毒素（BONT A）是人类最有效的毒素之一，并且已成为生物战中的迫在眉睫的威胁。高亲和力中和抗体作为暴露前或暴露后预防，可以潜在地保护疾病肉毒杆菌。人类的单链可变碎片抗体（SCFV）以前已经鉴定出了BONT A，包括一种几乎可以完全中和Bont A的抗体。尽管极为有前途，但这些抗体对BontA的亲和力将是有益的。这将通过使SCFV与亲和力成熟度的成熟度相结合，从而实现SCFV与SCFV的相结合方法。然后，最有希望的抗体将转换为更稳定的格式，以进一步表征。抗体预防BONT A的细胞毒性作用的能力将由基于体外细胞的测定法确定。另外，将使用biacore研究这些抗体的结合亲和力和特异性。抗体键的缔合速率常数，解离速率常数和平衡解离常数将获得进化抗体的结合。