克隆和从抗原碎片库中进行选择,以识别位.
1Department of Medicine, New York University Grossman School of Medicine, New York, New York 10016, USA gregg.silverman@nyulangone.org.
Cold Spring Harbor protocols
|November 7, 2025
概括
鉴定抗体-抗原结合部位的特征对于理解免疫反应和开发治疗方法至关重要. 这项研究改进了菌体显示方法,以有效地识别抗原上的最小抗体表位.
科学领域:
- 免疫学 免疫学 免疫学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 了解免疫反应需要对抗体-抗原相互作用进行分子级别的表征.
- 与微生物蛋白质结合的抗体可以中和毒素,并有助于病原体清除.
- 菌体显示是研究蛋白质与蛋白质相互作用的强大技术.
研究的目的:
- 描述一种优化的工作流程,用于识别和表征由抗体识别的抗原特异性表位.
- 改进确定抗体和抗原之间最小结合接触点的方法.
主要方法:
- 产生大型菌体显示库,随机克隆到pComb-Opti8菌体载体中的基因片段.
- 将库电解成细菌,然后恢复菌体颗粒,显示菌体外衣蛋白VIII (pVIII) 合的基因片段.
- 选择表达最小表位体的基体克隆,用于表位体识别的高通量DNA测序,以及对抗体-表位体结合的表征.
主要成果:
- 建立了一个工作流程,以优化抗体介导选择.
- 该工作流允许使用高通量测序快速识别编码感兴趣的表位的基因子区域.
- 开发了进一步表征抗体-表皮质结合相互作用的方法.
结论:
- 描述的工作流程有效地识别和表征抗原特异性表征.
- 这种方法提升了在分子水平上研究抗体-抗原相互作用的研究.
- 精细的菌体显示方法有助于理解免疫反应和开发向治疗方法.
更多相关视频
08:09Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
Published on: March 24, 2017
9.9K
09:07Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations
Published on: August 22, 2019
11.4K
相关概念视频
01:31Hybridoma Technology
13.4K
Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation,...
Hybridoma Selection
Commonly used fusion techniques — electroporation,...
13.4K
01:20Immunoprecipitation
5.6K
Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
5.6K