STTR Phase I: A Non-Chromatographic Technique for Synthetic Oligodeoxynucleotide Purification

这一小型企业技术转让(STTR)项目的更广泛影响/商业潜力将是开发一种新的合成寡核苷酸纯化方法。目前，大多数寡聚脱氧核苷酸都是通过层析纯化的。这些技术昂贵或难以放大，不适合并行提纯多个不同的样品。该工艺易于规模化提纯，适合于并行提纯。需要合成寡核苷酸的几个领域将受益于该技术，包括用于合成生物学应用的基因组组装的寡脱氧核苷酸疗法和寡脱氧核苷酸。对于治疗性制造，拟议的技术预计将降低生产成本。对于合成生物学来说，瓶颈在于基因组的从头构建，这需要大量的合成寡核苷酸。使用拟议技术的并行提纯将使这些材料更负担得起。此外，所提出的技术可以很容易地扩展到提纯其他生物低聚物，包括多肽和寡糖。这一延期将对生物医学研究等领域产生很大影响。本项目一期工程提出了一种基于聚合捕捉法的人工合成寡核苷酸的纯化方法，用于人工合成的寡核苷酸的纯化。目前，大多数人工合成的寡核苷酸是通过层析方法纯化的，这种方法依赖于用溶剂洗脱时产物和杂质在固体基质中移动的速度之差。缺点包括昂贵的仪器设备、密集的劳动强度、大量有害溶剂的使用以及无法净化长序列。这种方法放大成本高，不适合并行提纯。本项目旨在通过聚合寡核苷酸纯化技术将捕获的全长序列商业化，以解决这些问题。该方法的工作原理是选择性地将可聚合基团标记到寡聚脱氧核苷酸产物上，将其聚合成不溶于水的聚合物，洗掉所有杂质，然后将产物从聚合物中裂解出来。由于分离产物和杂质的原理与层析方法有很大的不同，该技术具有许多优点，包括不需要昂贵的仪器，使用简单，废品比低，适合于长序列的纯化。此外，该新技术易于扩展到大规模纯化，并且可以很容易地用于并行纯化。

The broader impact/commercial potential of this Small Business Technology Transfer (STTR) project will be the development of a novel method for synthetic oligodeoxynucleotide purification. Currently, most oligodeoxynucleotides are purified using chromatography. The techniques are expensive or difficult to scale up, and unsuitable for parallel purification of multiple different samples. The proposed technology is easy to scale up for large-scale purification and suitable for parallel purification. Several areas that require synthetic oligodeoxynucleotides will benefit from the technology including oligodeoxynucleotide therapeutics and oligodeoxynucleotides used in genome assembly for synthetic biology applications. For therapeutic manufacturing, the proposed technology is expected to bring down the cost of production. For synthetic biology, the bottleneck is in the area is de novo construction of genomes, which requires large numbers of synthetic oligodeoxynucleotides. Parallel purification using the proposed technology will make these materials more affordable. In addition, the proposed technology can be readily extended to purify other biooligomers including peptides and oligosaccharides. This extension will have a high impact in areas such as biomedical research. This STTR Phase I project proposes to method for synthetic oligodeoxynucleotide purification based on the "catching by polymerization" concept for the purification of synthetic oligodeoxynucleotides. Currently, most synthetic oligodeoxynucleotides are purified using chromatography methods, which rely on the rate of speed difference at which product and impurities travel in a solid matrix when eluted with solvents for separation. Drawbacks include expensive instrumentation, intensive labor, use of large volumes of harmful solvents and inability to purify long sequences. This method is expensive to scale up and unsuitable for parallel purification. This project aims to commercialize the catching full-length sequence by polymerization oligodeoxynucleotide purification technology to solve these problems. The method works by selectively tagging a polymerizable group to the oligodeoxynucleotide product, polymerizing it into an insoluble polymer, washing away all impurities and then cleaving the product from the polymer. Because the principle on which the product is separated from impurities is drastically different from that of chromatography methods, the proposed technique has many advantages, which include no need for expensive instrumentation, simple-to-use, low waste to product ratio, and suitability for purification of long sequences. Additionally, the new technique is readily scalable for large-scale purification, and can be easily adopted for parallel purification.