STTR Phase I: Purification of Mycosporine-like Amino Acids using Aptamers

STTR 第一阶段：使用适体纯化类菌孢菌素氨基酸

• 批准号: 1940352
• 负责人: David Smernoff
• 金额: $ 22.5万
• 依托单位: Heliobiosys, Inc.
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1940352&HistoricalAwards=false
• 关键词: STTR; Phase; Purification; Mycosporine; like

The broader impact of this STTR Phase I project is to harvest a natural sunscreen for broader distribution. This project will use a group of fast-growing microorganisms that make these ingredients. The challenge will be to harvest enough of the material to make effective and affordable sunscreens. The problem is akin to trying to collect all the "needles in a haystack". New tools that incorporate reusable analogues to "magnets" for capturing and releasing the sunscreen compound will provide a novel way to get the material out of a complex mixture of broken open cells and into the sunscreens. There is a lack of genuinely non-toxic, environmentally safe sunscreens in the U.S. market today. Melanoma, the most common cancer in the U.S., is a deadly form of skin cancer directly attributable to sun damage to skin, and conventional sunscreens are under pressure because of human health concerns and possible impacts on coral reefs. This work will provide a new class of sunscreens safer for people and marine life. Unlocking the potential of this nature-designed sunscreen will improve human health and the environment.This STTR Phase I project will evaluate the technical feasibility of using aptamers (small nucleic acids) to isolate mycosporine-like amino acids (MAAs) from a complex mixture of compounds derived from cyanobacteria. Small molecules present challenges for aptamer development because they can be: difficult to immobilize, have multiple three-dimensional conformations, and few chemically distinguishing features. MAAs present unique challenges due to the similarity of their chemical structure with other cell lysate materials and wide variety of amino acid substitutions. This innovation entails the isolation and identification of the specific MAAs being produced, isolation of aptamers that are highly specific, and assessing process scalability. MAAs are not commercially available so reference standards will be derived from the culture broth. These materials will be utilized as targets in aptamer selection using a proprietary single-stranded DNA library of aptamer sequences to identify suitable candidates by next-generation sequencing and bioinformatics analysis. Those aptamer candidates will then be screened and evaluated using an affinity purification workflow and absorbance-based microplate assay. The end result of this work will be to establish if this is a robust and scalable affinity purification method for MAAs.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.

该STTR I期项目的更广泛的影响是收集天然防晒霜以进行更广泛的分布。 该项目将使用制造这些成分的一组快速增长的微生物。面临的挑战将是收获足够的材料以制造有效且负担得起的防晒霜。问题类似于试图收集所有“针织中的针头”。将可重复使用的类似物包含到“磁铁”以捕获和释放防晒霜的新工具将提供一种新颖的方法，使材料从破碎的开放式细胞的复杂混合物中脱离并进入防晒霜。当今美国市场缺乏真正无毒的环境安全防晒霜。黑色素瘤是美国最常见的癌症，是一种致命的皮肤癌的形式，直接归因于阳光对皮肤的损害，而传统的防晒霜由于人类健康的关注以及对珊瑚礁的可能影响而受到压力。这项工作将为人们和海洋生物提供新的防晒霜。释放这种自然设计的防晒霜的潜力将改善人类健康和环境。该STTR I期项目将评估使用适体（小核酸）分离出从氰基果实中得出的复杂化合物的混合物的技术可行性。小分子对适体发育提出了挑战，因为它们可能是：难以固定，具有多个三维构象，几乎没有化学区分的特征。 MAA由于化学结构与其他细胞裂解物材料和各种氨基酸取代的相似性而提出了独特的挑战。这项创新需要隔离和识别所产生的特定MAA，隔离高度特异性的适体，并评估过程可伸缩性。 MAAS无法商业可用，因此参考标准将来自文化汤。 这些材料将使用适体序列的专有单链DNA库中的适体选择中用作目标，以通过下一代测序和生物信息学分析来识别合适的候选者。然后，将使用亲和力纯化工作流和基于吸光度的微板测定法对这些适体候选者进行筛选和评估。这项工作的最终结果将是确定这是否是MAAS的强大且可扩展的亲和力纯化方法。该奖项反映了NSF的法定任务，并且使用基金会的知识分子优点和更广泛的审查标准，被认为是值得通过评估来支持的。