Collaborative Research: Cell-free glycoprotein synthesis technology for point-of-care vaccine biomanufacturing

合作研究：用于即时疫苗生物制造的无细胞糖蛋白合成技术

• 批准号: 1936789
• 负责人: Michael Jewett
• 金额: $ 39.03万
• 依托单位: Northwestern University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-15 至 2023-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1936789&HistoricalAwards=false
• 关键词: Collaborative; Research; Cell; free; glycoprotein

Drug-resistant bacteria are a growing threat to human health. By the year 2050, up to 10 million lives per year could be at risk. New strategies will be needed to counter this threat. Vaccines have been developed to safely and effectively prevent dangerous bacterial infections. This project seeks to address current limitations in vaccine production. Cell-free technology for vaccine production that can be easily scaled up will be developed. This could lead to portable, on-demand vaccine development and production. The project will advance the biomanufacturing of conjugate vaccines. This would improve their availability to resource-poor communities. In parallel, hands-on learning modules will be developed and delivered to underrepresented high school students, and to undergraduate and graduate students. The bacterial cell surface is decorated with complex sugar structures. These include capsular polysaccharides (CPS) and O-polysaccharides (O-PS). These structures are important virulence factors, adhesion mediators, and immunomodulators. CPS and O-PS structures are typically absent from the surfaces of host cells. These polysaccharides can, therefore, be formulated as vaccine subunits and used to protect humans against life-threatening bacterial infections. Conjugate vaccines are a type of subunit vaccine whereby polysaccharide antigens are linked to a protein carrier. They are among the safest and most effective methods for inducing immunity against pathogenic bacteria. Current production technology is technically complex and relies on living cells. Refrigeration is necessary at every step along the distribution chain. As a result, manufacturing is centralized, capital intensive, and requires highly skilled labor. This project will create a scalable, cell-free biosynthesis technology for generating conjugate vaccine candidates. Experimental and computational approaches will be combined to develop and optimize a one-pot cell-free glycoprotein synthesis system. These glycoconjugates can be stored in freeze-dried formats and reconstituted simply by adding water. A complementary technology, shotgun scanning glycomutagenesis, will enable comprehensive evaluation of glycosylation efficiency as a function of conjugation site and antigen loading density, which will be correlated with vaccine immunogenicity.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

耐药细菌是对人类健康的日益严重威胁。到2050年，每年最多1000万生命可能会面临风险。应对这种威胁需要新的策略。已经开发出疫苗是为了安全有效地预防危险的细菌感染。该项目旨在解决疫苗生产中的当前局限性。可以开发可轻松扩展疫苗生产的无细胞技术。这可能会导致便携式，点播疫苗的开发和生产。该项目将推进结合疫苗的生物制造。这将提高他们对资源贫困社区的可用性。同时，动手学习模块将被开发并交付给占代表性不足的高中生，以及本科生和研究生。细菌细胞表面装饰有复杂的糖结构。这些包括囊膜多糖（CPS）和O-杆菌（O-PS）。这些结构是重要的毒力因子，粘附介质和免疫调节剂。 CPS和O-PS结构通常从宿主细胞的表面不存在。因此，这些多糖可以作为疫苗亚基配制，并用于保护人类免受威胁生命的细菌感染。结合疫苗是一种亚基疫苗，从而将多糖抗原与蛋白质载体相关。它们是诱导对病原细菌免疫的最安全，最有效的方法之一。当前的生产技术在技术上很复杂，依赖于活细胞。沿分销链的每个步骤都必须制冷。结果，制造业是集中的，资本密集的，需要高技能的劳动力。该项目将创建一种可扩展的，无细胞的生物合成技术，用于生成共轭疫苗候选物。实验和计算方法将合并，以开发和优化一个无细胞的糖蛋白合成系统。这些糖缀合物可以以冷冻干燥的格式存储，并简单地通过加水来重组。互补的技术，shot弹式扫描糖果量，将对糖基化效率作为结合位点和抗原载荷密度的函数进行全面评估，这将与疫苗免疫原性相关。该奖项将反映出NSF的法定任务，并通过评估范围来反映出构成的范围，该奖项已被视为众所周知的知识群体和富有的依据。

项目成果

Development of a Freeze-Dried CRISPR-Cas12 Sensor for Detecting Wolbachia in the Secondary Science Classroom

• DOI: 10.1021/acssynbio.1c00503
• 发表时间: 2022-02-18
• 期刊: ACS SYNTHETIC BIOLOGY
• 影响因子: 4.7
• 作者: Rybnicky, Grant A.;Dixon, Radeen A.;Jewett, Michael C.
• 通讯作者: Jewett, Michael C.

Point-of-Care Peptide Hormone Production Enabled by Cell-Free Protein Synthesis

通过无细胞蛋白质合成实现护理点肽激素生产

• DOI: 10.1021/acssynbio.2c00680
• 发表时间: 2023
• 期刊: ACS Synthetic Biology
• 影响因子: 4.7
• 作者: DeWinter, Madison A.;Thames, Ariel Helms;Guerrero, Laura;Kightlinger, Weston;Karim, Ashty S.;Jewett, Michael C.
• 通讯作者: Jewett, Michael C.

GlycoCAP: A Cell-Free, Bacterial Glycosylation Platform for Building Clickable Azido-Sialoglycoproteins

GlycoCAP：用于构建可点击叠氮基唾液酸糖蛋白的无细胞细菌糖基化平台

• DOI: 10.1021/acssynbio.3c00017
• 发表时间: 2023
• 期刊: ACS Synthetic Biology
• 影响因子: 4.7
• 作者: Thames, Ariel Helms;Moons, Sam J.;Wong, Derek A.;Boltje, Thomas J.;Bochner, Bruce S.;Jewett, Michael C.
• 通讯作者: Jewett, Michael C.

Cell-free expression and characterization of multivalent rhamnose-binding lectins using bio-layer interferometry

使用生物层干涉测量法进行多价鼠李糖结合凝集素的无细胞表达和表征

• DOI: 10.1093/glycob/cwad018
• 发表时间: 2023
• 期刊: Glycobiology
• 影响因子: 4.3
• 作者: Warfel, Katherine F.;Laigre, Eugénie;Sobol, Sarah E.;Gillon, Emilie;Varrot, Annabelle;Renaudet, Olivier;Dejeu, Jerome;Jewett, Michael C.;Imberty, Anne
• 通讯作者: Imberty, Anne

A universal glycoenzyme biosynthesis pipeline that enables efficient cell-free remodeling of glycans.

• DOI: 10.1038/s41467-022-34029-7
• 发表时间: 2022-10-24
• 期刊: Nature communications
• 影响因子: 16.6