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

这个STTR第一阶段项目的更广泛的影响是收获一种天然防晒霜，用于更广泛的销售。 该项目将使用一组快速生长的微生物来制造这些成分。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的“大海捞针”。新的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化合物，这将提供一种新的方法，将材料从破碎的开放细胞的复杂混合物中提取出来，并进入防晒霜。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损害，传统的防晒霜由于对人类健康的担忧和对珊瑚礁的可能影响而面临压力。这项工作将提供一种对人类和海洋生物更安全的新型防晒霜。释放这种天然防晒霜的潜力将改善人类健康和环境。STTR第一阶段项目将评估使用适体（小核酸）从蓝藻衍生化合物的复杂混合物中分离类菌孢素氨基酸（MAA）的技术可行性。小分子对适体开发提出了挑战，因为它们可能：难以降解，具有多种三维构象，并且几乎没有化学区别特征。由于其化学结构与其他细胞裂解物材料的相似性和各种氨基酸取代，MAA存在独特的挑战。这一创新需要分离和鉴定所产生的特定MAA，分离高度特异性的适体，并评估工艺可扩展性。MAA未市售，因此参比标准品将来自培养液。 这些材料将用作适体选择中的靶标，使用适体序列的专有单链DNA文库，通过下一代测序和生物信息学分析鉴定合适的候选物。然后使用亲和纯化工作流程和基于吸光度的微孔板测定法筛选和评价这些适体候选物。这项工作的最终结果将是确定这是否是一个强大的和可扩展的MAAs亲和纯化方法。该奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

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.