The challenge to develop a high throughput platform method for virus-based therapeutics has been stimulating the biomanufacturing community for decades. The potential of alternate technologies that can meet the current requirements might be better than the conventional methods for larger and more complex yet fragile moieties. One of those candidates is an aqueous two-phase system (ATPS), formed by two partially miscible aqueous solutions. ATPS are known for being inexpensive, mild, environmentally-friendly, and can be easily scaled-up and processed in a continuous mode. However, the challenge of not fully understanding the partitioning mechanism had resulted in a large experimental setup to determine optimal compositions specific to different viruses. Our DOE using a tie-line framework and study of virus surface characteristics provides a method to discern the interaction between virus and phase compositions. The current experimental design in a polyethylene glycol (PEG)-citrate system at physiological pH with the porcine parvovirus (PPV, a non-enveloped virus) facilitated in achieving a high yield of 80% in the PEG-rich phase. The resulting preferential partitioning supported the hypothesis of major influence of hydrophobic and electrostatic (salting-out) interactions between virus and relative phases. The framework was then utilized to create a database for five more viruses having different characteristics of presence of envelope, pI, and size. This multivariate data is being employed to empirically predict and reduce experimental space. This work demonstrates the purification efficacy of herpes simplex virus (HSV-1), pseudorabies virus (SuHV-1), human rhinovirus (HRV-14), human immunodeficiency virus-like particle (HIV-gag VLP), MS2 bacteriophage and ways to generalize optimization of target virus in ATPS. The ATPS approach will not only help generalize virus purification but also provide a high throughput technology in virus-based therapeutics DSP.

Generalized purification of viruses and virus-like particles using aqueous two-phase system

Macromolecular Chemistry: The Second Century

[Author Benefits](https://scimeetings.acs.org/?utm_source=pubs_content_marketing&utm_medium=website&utm_campaign=0320_MCF_NPI_Launch_Spring_Homepage&ref=pubs_content_marketing)

[How to use SciMeetings](https://storage.googleapis.com/dl.morressier.com/acs/SciMeetings-ACSSpring2020-HowToGuide_M3020-Final.pdf)

[Frequently Asked Questions](https://storage.googleapis.com/dl.morressier.com/acs/SciMtng_ACSNMSp2020_FAQ_M3120_Final.pdf)

ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.