Characterization of therapeutic human monoclonal antibodies against SARS

抗 SARS 治疗性人单克隆抗体的表征

• 批准号: 7645228
• 负责人: Bellur S Prabhakar
• 金额: $ 56.92万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-12 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7645228
• 关键词: Acute; Affect; Affinity; Amino Acid Sequence; Amino Acid Substitution; Animals; Antibodies; Antigens; Area; Back; Base Sequence; Binding; Biological Assay; Cell fusion; Cell-Matrix Junction; Clinical; Collaborations; Data; Disease; Dose; Epitopes; Escape Mutant; Gene Expression; Genes; Genomics; Glycoproteins; Human; Hybridomas; Immune response; Immunotherapeutic agent; In Vitro; Infection; Mammalian Cell; Mediating; Modality; Monitor; Monoclonal Antibodies; Mutation; Oligonucleotides; Passive Immunization; Patients; Peptides; Prevention; Prophylactic treatment; Protein Fragment; Proteins; Proteolysis; Recombinants; Relative (related person); SARS coronavirus; Serum; Severe Acute Respiratory Syndrome; Specificity; Surface; Technology; Testing; Text; Therapeutic; TimeLine; Transfection; Vaccinated; Variant; Viral; Virus; Virus Diseases; Western Blotting; base; glycosylation; human monoclonal antibodies; human subject; in vivo; interest; mortality; mutant; neutralizing antibody; novel; prevent; protein expression; prototype; receptor binding; research study

Characterization of Therapeutic human monoclonal antibodies against SARS The discovery that a corona virus is the causative agent of SARS and its rapid genomic sequencing has created opportunities to develop effective modalities of prevention and treatment against this virus. Of particular interest is the spike glycoprotein (S protein), which is expressed as a surface projection. The S protein is involved in viral attachment, cell-cell fusion, the induction of protective immune response in virus neutralization. This proposal will focus on developing human monoclonal antibodies that can be used for prophylaxis, and/or post-exposure treatment of SARS-CoV infection. We have already generated a panel of hybridomas capable of producing neutralizing human monoclonal antibodies (HmAbs) in close collaboration with Abgenix Inc using their XenoMouse. These hybridomas have been cloned and 19 HmAbs have been produced, purified and re-tested in a neutralization assay. In addition, we have already produced several recombinant fragments of the S protein and obtained overlapping peptides that span the S protein, and used them to determine binding specificities of HmAbs. In aim-1 we will further characterize the neutralizing ability of human monoclonal antibodies against SARS-CoV in vitro and determine their relative binding affinities and specificities using a number of different current approaches. In aim-2, we will identify human monoclonal antibodies that can potentially neutralize a broad range of clinical isolates by testing them in a viral entry and a cell fusion assay using different pseudotyped viruses expressing mutant S proteins that contain amino acid substitutions identified in over 90 clinical isolates from around the world. These studies are expected to result in the characterization of HmAbs with potential as passive immunotherapeutics against SARS-CoV infection in humans.

抗 SARS 治疗性人单克隆抗体的表征 冠状病毒是 SARS 病原体的发现及其快速基因组测序为开发针对该病毒的有效预防和治疗方式创造了机会。特别令人感兴趣的是刺突糖蛋白（S 蛋白），它表示为表面投影。 S蛋白参与病毒附着、细胞间融合、病毒保护性免疫反应的诱导 中和。该提案将重点开发可用于预防和/或暴露后治疗 SARS-CoV 感染的人类单克隆抗体。我们已经与 Abgenix Inc 密切合作，使用他们的 XenoMouse 生成了一组能够产生中和人单克隆抗体 (HmAb) 的杂交瘤。这些杂交瘤已被克隆，19 种 HmAb 已被生产、纯化并在中和测定中重新测试。此外，我们已经生产了几个S蛋白的重组片段，并获得了跨越S蛋白的重叠肽，并用它们来确定HmAb的结合特异性。在目标 1 中，我们将进一步表征人类单克隆抗体在体外对抗 SARS-CoV 的中和能力，并使用多种不同的当前方法确定它们的相对结合亲和力和特异性。在目标 2 中，我们将鉴定出可以中和多种抗体的人类单克隆抗体。 通过使用表达突变 S 蛋白的不同假型病毒进行病毒进入和细胞融合测定，对临床分离株进行测试，这些突变 S 蛋白含有在来自世界各地的 90 多个临床分离株中发现的氨基酸取代。这些研究预计将得出 HmAb 的特征，这些 HmAb 有可能作为人类 SARS-CoV 感染的被动免疫疗法。