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

这个STTR第一期项目的更广泛影响是收获一种天然防晒霜，用于更广泛的分布。这个项目将使用一组快速生长的微生物来制造这些成分。面临的挑战将是收获足够的材料来制造有效和负担得起的防晒霜。这个问题类似于试图收集所有“干草堆里的针”。新工具将可重复使用的类似物与“磁铁”结合起来，用于捕获和释放防晒霜化合物，这将提供一种新方法，将材料从破碎的开放细胞的复杂混合物中提取出来，进入防晒霜。目前，美国市场上缺乏真正无毒、环保的防晒霜。黑色素瘤是美国最常见的癌症，是一种致命的皮肤癌，直接归因于阳光对皮肤的伤害，由于人类健康问题和可能对珊瑚礁的影响，传统的防晒霜正面临压力。这项工作将为人类和海洋生物提供一种更安全的新型防晒霜。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一期项目将评估使用适体（小核酸）从蓝藻菌衍生的复杂化合物混合物中分离真菌孢素样氨基酸（MAAs）的技术可行性。小分子对适体的发展提出了挑战，因为它们难以固定，具有多个三维构象，并且几乎没有化学特征。MAAs由于其化学结构与其他细胞裂解物材料的相似性和广泛的氨基酸取代而面临独特的挑战。这种创新需要隔离和识别正在生产的特定MAAs，隔离高度特定的适配体，并评估过程的可扩展性。MAAs不是市售的，所以参考标准将从培养液中得到。这些材料将用作适体选择的靶标，使用专有的适体序列单链DNA文库，通过下一代测序和生物信息学分析确定合适的候选体。然后使用亲和纯化工作流程和基于吸光度的微孔板检测筛选和评估这些适体候选体。这项工作的最终结果将是确定这是否是一种健壮且可扩展的maa亲和纯化方法。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.