Structural, thermodynamic and kinetic studies of antibo*

抗体的结构、热力学和动力学研究*

• 批准号: 6402341
• 负责人: BRADFORD C BRADEN
• 金额: $ 3.74万
• 依托单位: BOWIE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6402341
• 关键词: X ray crystallography; antiantibody; antigen antibody reaction; antiidiotype antibody; binding sites; cell component structure /function; chemical kinetics; cooperative study; fluorescence polarization; hybridomas; immunoglobulin G; immunoglobulin M; immunoglobulins; intermolecular interaction; laboratory mouse; lysozyme; microcalorimetry; molecular cloning; monoclonal antibody; nucleic acid sequence; protein binding; stop flow technique; structural biology; surface plasmon resonance; thermodynamics; ultracentrifugation

DESCRIPTION (provided by applicant) Affinity maturation of the immune response is a process unique to the immune system. This process consists of somatic hypermutation of the variable domains of immunoglobulins, which leads to a significant increase in the affinities of the antibodies elicited during a humoral immune response. The understanding of the molecular and structural basis of this phenomenon is an area of particular interest for Immunology as well as the biological sciences in general. Several authors have described, in detail, models of affinity maturation in antibody-hapten systems. However, affinity maturation against protein antigens has not been described in such detail. Our previous work, as in other laboratories, has shown that the antibodies elicited during the immune response against a protein antigen show a fast maturation of the affinity, a phenomenon closely related to the early IgM-IgG isotype class switch process. No structural, sequence or functional studies describing the changes that occur at the combining site of anti-protein antibodies during this process can be found in the current literature. The aim of this project is to characterize the sequence and the binding site structures of the IgM unmutated precursors of the structurally well characterized IgG anti-hen egg-white lysozyme (HEL) antibodies. This goal will be accomplished by cloning and sequencing anti-HEL IgMs and by X-ray crystallographic structure determinations of the Fv fragments of these IgMs. Structural, functional and mutational studies of the recombinant Fv fragments of those IgMs would reveal the nature of the changes produced during affinity maturation.

描述（由申请人提供） 免疫应答的亲和力成熟是免疫应答特有的过程。 系统这个过程包括可变结构域的体细胞超突变 免疫球蛋白，这导致显着增加的亲和力， 体液免疫反应中产生的抗体的理解 这种现象的分子和结构基础是一个特别的领域， 对免疫学和一般生物科学感兴趣。 几位作者详细描述了亲和力成熟的模型， 抗体-半抗原系统。然而，针对蛋白质抗原的亲和力成熟 还没有被如此详细地描述。我们以前的工作，如在其他 实验室，已经表明，在免疫反应过程中引发的抗体 针对蛋白质抗原显示出快速成熟的亲和力，这是一种现象， 与早期IgM-IgG同种型类别转换过程密切相关。没有 结构、序列或功能研究，描述发生在 在此过程中，可以找到抗蛋白抗体的结合位点 在目前的文学。 本项目的目的是表征序列和结合位点 结构良好的IgM未突变前体的结构 特征性IgG抗鸡蛋清溶菌酶（HEL）抗体。这一目标将 通过克隆和测序抗HEL IgM和通过X射线 这些IgM的Fv片段的晶体结构测定。 重组Fv片段的结构、功能和突变研究 这些IgM将揭示亲和过程中产生的变化的性质， 成熟