Anything but conventional chromatography, also known as the ABC trend, is currently attracting interest to develop purification alternatives to packed-bed chromatography. Precipitation and magnetic separation are two techniques already proven to accomplish protein purification from complex media alone, yet never used in synergy.<br/>With the aim to capture antibodies directly from crude extracts, a new method combining precipitation and magnetic separation was developed and coined affinity magnetic precipitation. Precipitation screens based on the Hofmeister series and a commercial precipitation kit, were tested with magnetic particles for antibody capture from different sources.<br/>Two different antibodies sources were tested: (i) polyclonal antibodies from human serum plasma, and (ii) anti-TNFα mAb from clarified CHO cell supernatants. The study comprised the discovery of the best precipitant and precipitation conditions using magnetic particles functionalized with a synthetic ligand previously developed. The addition of particles was crucial to achieve higher antibody purity and recovery. Nonetheless, the optimization of the precipitant and precipitation conditions also revealed to be essential to minimize antibody loss and maximize its purity. The elution was always performed using PBS at pH 7.4. As such, this approach may lead not only to antibody capture but also conditioning and concentration in one single step. Contrary to Protein A affinity chromatography, in our studied system, the elution conditions are mild. The activity of the eluted mAb was assessed In terms of process performance, the hybrid system enabled 80% purity and 50% recovery of polyclonal antibodies from plasma, and 97% recovery yield and 99% purity from anti-TNFα mAb. The presence of aggregates and the biological activity of the eluted fractions were evaluated, showing homogeneous eluted samples with high biological activity.<br/>The potential to use a hybrid system based on precipitation and magnetic fishing with affinity towards protein purification enables its selective capture and concentration in a single step unit operation.

Affinity magnetic precipitation for antibody purification

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.