Molecular and Structural Studies of Antibody Diversity Mechanisms

抗体多样性机制的分子和结构研究

• 批准号: 9319776
• 负责人: Vaughn Vasil Smider
• 金额: $ 36.58万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9319776
• 关键词: Affinity; Amino Acids; Antibodies; Antibody Diversity; Antigens; Architecture; Binding; Cattle; Color; Crystallization; Crystallography; Data; Diagnostic; Disulfides; Engineering; Flow Cytometry; Genetic; Human; Immunoglobulin Domain; Immunoglobulins; Knowledge; Lead; Libraries; Mammals; Medical; Modeling; Molecular; Mutagenesis; Pattern; Peptides; Pharmaceutical Preparations; Phylogeny; Process; Property; Protein Engineering; Proteins; Research; Role; Structure; Surface Immunoglobulins; System; Therapeutic; Variant; antibody engineering; antigen binding; deep sequencing; disulfide bond; drug development; experimental study; novel; physical property; public health relevance; scaffold

DESCRIPTION (provided by applicant): Antibody molecules are enormously important as therapeutic and diagnostic molecules. More recently, unique scaffolds like the VHH of camelids, or even non-antibody frameworks like "knottins" have become important in biomedicine. Antibody diversity and antigen binding in mammals is often restricted to the CDR loops of the immunoglobulin fold. In experiments challenging this paradigm, we have recently solved the crystal structures of two bovine antibodies containing ultralong CDR H3s (56 and 61 amino acids) and also deep sequenced the ultralong repertoire. Our data reveal that these CDR H3s form a very unusual architecture composed of a long �-strand "stalk" which supports a disulfide rich "knob" that protrudes far from the immunoglobulin surface. Interestingly, the two different antibodies contain different patterns of disulfides, which result in different knob structures. Dee sequencing reveals extensive diversity in the ultralong CDR H3s where a multitude of different disulfides could potentially form within the knob. Thus, the bovine antibody system can produce an unprecedented repertoire of mega CDR H3s that may result in an impressive diversity of minifolds containing combinations of somatically generated disulfides. Thus, antibody diversity is located in a new minifold supported by the immunoglobulin domain. We will perform structural, functional, and engineering studies to investigate the properties of this new antibody class, as well as to lead the way to developing this unique structure into therapeutics.

描述（由适用提供）：抗体分子作为治疗和诊断分子非常重要。最近，诸如骆驼的VHH，甚至是“ Knottins”这样的非抗体框架等独特的脚手架在生物医学中变得很重要。哺乳动物中的抗体多样性和抗原结合通常仅限于免疫球蛋白折叠的CDR环。在实验挑战这一范式中，我们最近求解了两种含有超大CDR H3S（56和61个氨基酸）的牛抗体的晶体结构，并深入测序了Ultralong曲目。我们的数据表明，这些CDR H3S形成了一个非常不寻常的结构，该结构由长的“茎”组成，该结构支持富含二硫化物的“旋钮”，远离免疫球蛋白表面。有趣的是，两种含有不同二硫化物模式的不同抗体，从而导致不同的旋钮结构。 DEE测序揭示了超大的CDR H3中广泛的多样性，其中许多不同的二硫化物可能会在旋钮内形成。这是牛抗体系统可以产生巨型CDR H3S的前所未有的曲目，这可能会导致令人印象深刻的微型含量，这些微型含有含有体外产生的二硫化物组合的微型。这是抗体多样性位于由免疫球蛋白结构域支持的新的微型iFold中。我们将进行结构，功能和工程研究，以研究这种新抗体类别的特性，并为将这种独特的结构开发为治疗。