Aptamer-mediated purification for viruses

我的研究小组最近开发了可转换适体（SwAps），单链DNA寡核苷酸，对溶瘤病毒具有受控的亲和力和选择性，可以在用简单的通用洗脱剂处理后释放完整的病毒。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的水泡性口炎病毒VSV，它对难以通过常规疗法治疗的转移性癌症特别有效。没有其他VSV纯化方法接近我们的基于Swap的纯化技术的效率和简单性。早期的实验室研究已经最终表明，开发的SwAps可以在Ca和Mg离子的存在下有效地捕获VSV，并且还使用具有螯合剂如EDTA和EGTA的洗脱溶液释放捕获的病毒。我们现在正处于SwAps技术准备进入简化到实践阶段的阶段。该I2 I提案的第1阶段的总体目标是通过实现以下步骤将我们的基于SwAps的纯化技术发展成稳健且易于制造的平台技术：1）增强靶生物分子捕获和回收; 2）在频繁纯化循环期间改善SwAps稳定性; 3）研究经济且有效的色谱基质; 4）研究基于SwAps的纯化技术。4）纯化工艺的优化; 5）第三方评估和验证。这些目标的实现将导致开发一种强大的技术，该技术将满足参与制药和医学意义的生物分子纯化的客户的关键需求，并将为初创公司和可在短期内部署的产品线奠定基础。

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.