Expressing a novel class of heavy chain antibodies

表达一类新型重链抗体

• 批准号: 10432883
• 负责人: ELLEN HSU
• 金额: $ 20.19万
• 依托单位: SUNY DOWNSTATE MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-25 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10432883
• 关键词: Amino Acids; Antibodies; Antibody Repertoire; Antigens; Arginine; Bacterial Antigens; Bacterial Capsules; Binding; Binding Sites; Bypass; Cell Wall; Charge; Chondrichthyes; Complementary DNA; Coupled; Disease; Epitopes; Event; Exons; Fishes; Future; Genes; Haptens; Human; Immune response; Immunoglobulins; Immunotherapy; Infection; Ligand Binding; Ligands; Light; Llama; Lysine; Microbe; Molecular; Monoclonal Antibodies; Mormyridae; Mouse Cell Line; Mutant Strains Mice; Proteins; Reading Frames; Reagent; Reporting; Role; Shark; Streptococcus pneumoniae; Surface; Testing; Transgenes; Transgenic Mice; V(D)J Recombination; Vertebrates; antibody detection; antigen binding; base; drug development; experimental study; flexibility; novel; pathogen; polypeptide; tool

PROJECT SUMMARY   Heavy chain antibodies (HCAbs) are an unusual species of immunoglobulins expressed by camelids and sharks. They occur as homodimers in which the variable (V) domain of each polypeptide binds ligand. Without the requirement for the light chain partner in conventional antibodies, HCAb reagents have the advantage of size and molecular manipulability and are promising tools for drug development and immunotherapy. Genes encoding HCAbs evolved separately in two subclasses of cartilaginous fish that diverged 420 million years ago. The role of their HCAbs and pathogen recognition capabilities have not been characterized, although it has been determined that HCAb convex antigen-combining sites can detect epitopes inaccessible or bypassed by conventional antibodies. We have discovered novel antibody genes in a cartilaginous fish whose component gene segments have evolved to encode and rearrange to express lysines and arginines at the ligand-binding sites. An antibody repertoire whose major antigen-combining loop is flexible and electropositive is unique among vertebrates, presenting an opportunity to seek antigenic determinants different from what has been classically defined. Transgenic mice will be generated to express a set of chimeric fish genes. The first aim of this proposal is to generate the mutant mice, the second is to test the ability of the transgene repertoire to respond to various immunogens. Experiments are proposed to test for HCAb detection of novel epitopes in bacterial capsule and cell wall. Future studies will ascertain their protectiveness against disease.

项目概要   重链抗体 (HCAb) 是骆驼科动物表达的一种不寻常的免疫球蛋白 和鲨鱼。它们以同二聚体的形式出现，其中每个多肽的可变 (V) 结构域结合配体。 HCAb 试剂不需要传统抗体中的轻链伴侣， 尺寸和分子可操作性的优势，是药物开发和药物开发的有前途的工具 免疫疗法。 第420章 编码HCAb的基因在软骨鱼的两个亚类中分别进化 万年前。它们的 HCAb 和病原体识别能力的作用尚未得到证实。 尽管已确定 HCAb 凸面抗原结合位点可以检测到 常规抗体无法接近或绕过的表位。我们发现了新的抗体基因 一种软骨鱼，其组成基因片段已进化为编码和重新排列以表达 配体结合位点上的赖氨酸和精氨酸。一种抗体库，其主要抗原结合 环是灵活的，正电性在脊椎动物中是独一无二的，提供了寻找抗原的机会 与经典定义不同的决定因素。 将产生转基因小鼠来表达一组嵌合鱼基因。本次活动的第一个目的 建议一是产生突变小鼠，二是测试转基因库的能力 对各种免疫原作出反应。建议进行实验来测试 HCAb 对新表位的检测 细菌荚膜和细胞壁。未来的研究将确定它们对疾病的保护作用。