Array Based Affinity Selection

基于阵列的亲和力选择

• 批准号: 10163532
• 负责人: BRIAN KENNETH KAY
• 金额: $ 18.33万
• 依托单位: TANGO BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10163532
• 关键词: 2019-nCoV; ARNT gene; Administrative Supplement; Affinity; Animals; Antibodies; Arizona; Award; Binding; Biological Assay; Biological Sciences; CDK2 gene; COPS5 gene; Cancer Etiology; Cells; Chimeric Proteins; Chinese Hamster Ovary Cell; Collaborations; Complex Mixtures; Contract Services; DNA Sequence Alteration; Diagnostic; Ensure; Family; Fibronectins; Funding; Generations; Genetic Diseases; Genetic Transcription; Glass; Goals; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Human; Human Cell Line; Human Engineering; Human Genome; Immunization; Immunologics; In Situ; In Vitro; Individual; Lateral; Libraries; Location; Malignant Neoplasms; Mass Spectrum Analysis; Methods; Modeling; Monomeric GTP-Binding Proteins; Mus; National Institute of General Medical Sciences; Oryctolagus cuniculus; Pathway interactions; Patients; Phage Display; Pharmacologic Substance; Phase; Plasmids; Process; Property; Protein Array; Protein Family; Proteins; Proteome; Protocols documentation; PubMed; Publications; Reagent; Recombinant Antibody; Recombinants; Recovery; Research; Resources; Route; Saliva; Scaffolding Protein; Slide; Small Business Technology Transfer Research; Specificity; Spottings; Technology; Therapeutic; Therapeutic Agents; Time; Tissues; Translating; Translations; Universities; Viral; Virion; Virus; Work; base; commercial application; diagnostic assay; experience; experimental study; member; method development; milligram; mutant; nanomolar; novel; overexpression; protein purification; receptor binding; screening; suicide substrates

Project Summary Funds are requested to apply technologies being developed in a current NIGMS award to generate recombinant antibody-like affinity reagents to the spike protein of the SARS-CoV-2 virus. As a proof-of-principle experiment, we have already isolated four fibronectin type III (FN3) monobodies that bind with low nanomolar affinity to the receptor binding domain (RBD) of the viral spike protein. An administrative supplement will permit continuation and expansion of this work to generate affinity reagents that bind with picomolar affinity and can be used in sensitive, robust diagnostic assays (homogenous assays, lateral flow assays) for virus particles in saliva. Separately, these monobodies will be reformatted as bivalent Fc fusions and overexpressed in CHO cells. Such reagents have the potential to serve as therapeutic agents that lower or block viral entry into patients ACE2-expressing cells. 1

项目概要 申请资金应用当前 NIGMS 奖项中正在开发的技术，以生成针对 SARS-CoV-2 病毒刺突蛋白的重组抗体样亲和试剂。作为一个 原理验证实验，我们已经分离出四种纤连蛋白 III 型 (FN3) 单体 以低纳摩尔亲和力与病毒刺突蛋白的受体结合域（RBD）结合。行政补充将允许这项工作继续和扩大，以产生 亲和试剂以皮摩尔亲和力结合，可用于灵敏、稳健的诊断 唾液中病毒颗粒的测定（同质测定、侧流测定）。分别地，这些 单体将被重新格式化为二价 Fc 融合体并在 CHO 细胞中过表达。这样的 该试剂有潜力作为治疗剂，降低或阻止病毒进入患者表达 ACE2 的细胞。 1

项目成果

FN3-based monobodies selective for the receptor binding domain of the SARS-CoV-2 spike protein.

SARS-COV-2尖峰蛋白的受体结合结构域选择性基于FN3的单体反应。

• DOI: 10.1016/j.nbt.2021.01.010
• 发表时间: 2021-05-25
• 期刊: New biotechnology
• 影响因子: 5.4
• 作者: Miller CJ;McGinnis JE;Martinez MJ;Wang G;Zhou J;Simmons E;Amet T;Abdeen SJ;Van Huysse JW;Bowsher RR;Kay BK
• 通讯作者: Kay BK