Expressing a novel class of heavy chain antibodies

表达一类新型重链抗体

• 批准号: 10555257
• 负责人: ELLEN HSU
• 金额: $ 24.23万
• 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 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10555257
• 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; 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.

项目摘要   重链抗体（HCAB）是骆驼表达的一种不寻常的免疫球蛋白物种 和鲨鱼。它们以同型二聚体的形式出现，其中每个多肽的变量（V）结构域结合配体。 无需在常规抗体中需要轻链伴侣，HCAB试剂具有 大小和分子可操作性的优势，是药物开发和 免疫疗法。 编码HCABS的基因分别演变在两个软骨鱼的子类中，分别有420 百万年前。他们的HCAB和病原体识别能力的作用尚未 表征，尽管已经确定HCAB凸抗抗原组合位点可以检测到 表位无法接近或被传统抗体绕开。我们在 一个软骨鱼的基因段的软骨鱼已经演变为编码和重新排列以表达 配体结合位点的赖氨酸和精氨酸。抗体曲目，其主要抗原组合 环具有灵活性，电阳性在脊椎动物中是独一无二的，这是一个寻求抗原的机会 确定与经典定义的不同。 转基因小鼠将产生以表达一组嵌合鱼基因。第一个目的 建议是生成突变小鼠，第二是测试转换曲目的能力 应对各种免疫原。提出了实验来测试HCAB在新表位的HCAB检测 细菌胶囊和细胞壁。未来的研究将确定它们对疾病的保护。