Recombinant Engineering of SARS-CoV-2 Spike and N proteins
SARS-CoV-2 刺突蛋白和 N 蛋白的重组工程
基本信息
- 批准号:10272263
- 负责人:
- 金额:$ 3.86万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:
- 资助国家:美国
- 起止时间:至
- 项目状态:未结题
- 来源:
- 关键词:2019-nCoVAffinityAngiotensinsAntibodiesBindingBioinformaticsComplexComputing MethodologiesEngineeringEscherichia coliExhibitsFc ReceptorGoalsLengthMutagenesisMutationNucleoproteinsPeptidyl-Dipeptidase AProtein EngineeringProteinsPublishingRecombinantsResolutionSeriesSiteStructureSurface Plasmon ResonanceSystemX-Ray Crystallographydesignexpression vectorimprovedmutantnanobodiesreceptorreceptor bindingscreening
项目摘要
Our goal in this project is to explore published structures of the SARS-CoV-2 Spike protein with a focus on its receptor binding domain (RBD), in efforts to generate recombinant molecules of increased thermal stability. To this end we have examined published structures and bioinformatically have made a number of predictions of site-directed mutants that may yield molecules of increased stability. Initial studies have confirmed our ability to produce the wild type RBD in good yield from E. coli expression vectors, to produce the molecule in high purity through multiple chromatographic steps, and to confirm its ability to bind specific antibodies. Mutational studies are in progress. In addition, we have obtained vectors for the expression of the full-length Spike protein and its lumens domain, and expression studies in mammalian systems are underway. Also, we are reengineering the angiotensin converting enzyme-2 (ACE-2) receptor for similar expression, binding, and mutagenesis studies.
我们在这个项目中的目标是探索已发表的SARS-CoV-2刺突蛋白的结构,重点是其受体结合结构域(RBD),努力产生热稳定性增加的重组分子。为此,我们已经研究了已发表的结构和生物信息学已经作出了一些预测的定点突变体,可能会产生分子的稳定性增加。初步的研究已经证实了我们从E.大肠杆菌表达载体,通过多个层析步骤生产高纯度的分子,并确认其结合特异性抗体的能力。突变研究正在进行中。此外,我们已经获得了用于表达全长Spike蛋白及其管腔结构域的载体,并且正在哺乳动物系统中进行表达研究。此外,我们正在重新设计血管紧张素转换酶-2(ACE-2)受体,用于类似的表达、结合和诱变研究。
项目成果
期刊论文数量(0)
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科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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David Margulies其他文献
David Margulies的其他文献
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{{ truncateString('David Margulies', 18)}}的其他基金
Variant detection and variant analysis process for diagnosis of CH and MODY
用于诊断 CH 和 MODY 的变异检测和变异分析流程
- 批准号:
7218897 - 财政年份:2006
- 资助金额:
$ 3.86万 - 项目类别:
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