Synthetic oligonucleotides contain impurities such as truncated failure sequences. Current methods for their purification include RP HPLC, IX HPLC, gel electrophoresis, biotin-avidin affinity purification, fluorous affinity purification and others. These methods are not ideal for certain applications due to shortcomings such as consumption of large volume of harmful solvents, high labor demand, expensive instrument and materials, and difficulty in scaling up and automation. We recently developed non-chromatographic purification methods based on the concept of catching-by-polymerization. In this method, the full-length sequences are tagged with an acrylation phosphoramidite. After synthesis, cleavage and deprotection, the crude ODN is subjected to radical polymerization. The full-length sequences are incorporated into polymer. After washing failure sequences and other impurities away, pure full-length sequences are cleaved from polymer. This method gives pure ODNs with good recovery yields. They do not need any type of chromatography. Purification is achieved by simple manipulations such as shaking and filtration. It is demonstrated that this method is useful for large-scale, as well as small-scale and high throughput purification and long ODN purification. Our work in the area has established a foundation to transform the entire multiple billion dollar industry of synthetic biooligomers

Synthetic oligodeoxynucleotide purification using the catching full-length sequence by polymerization technique

ACS Fall 2021

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