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

STTR 第一阶段：合成寡脱氧核苷酸纯化的非色谱技术

基本信息

批准号：
1720774
负责人：
Fengping Wei
金额：
$ 22.5万
依托单位：
CGeneTech, Inc.
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-01 至 2018-11-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1720774&HistoricalAwards=false
关键词：
STTR Phase Non Chromatographic Technique

项目摘要

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.

这项小型企业技术转让（STTR）项目的更广泛影响/商业潜力将是开发一种合成寡脱氧核苷酸纯化的新方法。目前，大多数寡脱氧核苷酸是用色谱法纯化的。该技术昂贵或难以扩大规模，并且不适合多个不同样品的平行纯化。该技术易于规模化提纯，适用于平行提纯。需要合成寡脱氧核苷酸的几个领域将受益于该技术，包括寡脱氧核苷酸治疗和用于合成生物学应用的基因组组装的寡脱氧核苷酸。对于治疗性生产，该技术有望降低生产成本。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脱氧核苷酸。采用所提出的技术进行平行净化将使这些材料更加经济实惠。此外，所提出的技术可以很容易地扩展到纯化其他生物低聚物，包括肽和低聚糖。这一扩展将在生物医学研究等领域产生重大影响。本STTR一期项目提出了一种基于“聚合捕集”纯化合成寡脱氧核苷酸概念的合成寡脱氧核苷酸纯化方法。目前，大多数合成的寡脱氧核苷酸都是用色谱法纯化的，这种方法依赖于用溶剂洗脱分离时产物和杂质在固体基质中移动的速度差率。缺点包括昂贵的仪器，密集的劳动，使用大量的有害溶剂和无法净化长序列。该方法规模扩大成本高，不适合平行净化。本项目旨在通过聚合低聚脱氧核苷酸纯化技术实现捕获全长序列的商业化。该方法通过选择性地将可聚合基团标记到寡脱氧核苷酸产物上，将其聚合成不溶性聚合物，洗去所有杂质，然后将产物从聚合物中分离出来。由于产品从杂质中分离的原理与色谱方法截然不同，因此所提出的技术具有许多优点，包括不需要昂贵的仪器，使用简单，废产物比低，适合纯化长序列。此外，新技术很容易扩展到大规模净化，并且可以很容易地采用平行净化。