Aptamer-mediated purification for viruses

我的研究小组最近开发了可切换适配体（swap），单链DNA寡核苷酸，对溶瘤病毒具有控制的亲和力和选择性，可以在简单的通用洗脱剂治疗后释放完整的病毒。这项新技术有可能彻底改变基于亲和力的纯化和分离。使用这种方法，我们能够获得一种超纯水疱性口炎病毒，VSV，它对传统疗法难以治疗的转移性癌症特别有效。没有其他的VSV净化方法可以接近我们基于swap的净化技术的效率和简单性。早期的实验室研究已经确凿地表明，开发的swap可以在Ca和Mg离子存在的情况下有效地捕获VSV，并使用含有螯合剂（如EDTA和EGTA）的洗脱液释放捕获的病毒。我们现在所处的阶段是，swap技术已经准备好进入简化到实践阶段。本I2I提案第一阶段的总体目标是通过实现以下步骤，将我们基于swap的纯化技术发展成为强大且易于制造的平台技术：1)增强目标生物分子的捕获和恢复；2)在频繁的净化循环中，swap的稳定性得到改善；3)经济高效色谱基质的研究；4)净化工艺的优化；5)第三方评估和验证。这些目标的实现将导致一项强大的技术的发展，该技术将满足涉及制药和医疗意义的生物分子纯化的客户的关键需求，并将形成可在短期内部署的初创公司和产品线的基础。

My research group recently developed Switchable Aptamers (SwAps), single-stranded DNA oligonucleotides, of controlled affinity and selectivity toward oncolytic viruses that can allow the release of the intact virus upon treatment with a simple universal eluting agent. This new technology has the potential to revolutionize affinity-based purification and separation. Using this method, we were able to obtain an ultrapure vesicular stomatitis virus, VSV, which is particularly effective against metastatic cancers that are difficult to treat via conventional therapy. There is no other method for VSV purification that approaches the efficiency and simplicity of our SwAp-based purification technique. Early laboratory studies have conclusively shown that the developed SwAps can efficiently capture VSV in the presence of Ca and Mg ions and also release the captured virus using an eluting solution with chelating agents such as EDTA and EGTA. We are now at a stage where the SwAps technology is ready to move to the reduction-to-practice stage. The overall goals for Phase 1 of this I2I proposal are to develop our SwAp-based purification technique into a robust and easily manufactured platform technology by achieving the following steps: 1) Enhancement of the target biomolecule capture and recovery; 2) Improvement of the SwAps stability during frequent purification cycles; 3) Investigation of economic and efficient chromatographic matrices; 4) Optimization of the purification process; 5) Third party evaluation and validation. Achievement of these goals will result in the development of a robust technology that will meet the key needs of customers involved in purification of biomolecules of pharmaceutical and medicinal significance and will form the basis for both a start-up company and a product line that can be deployed in short period.