AI Article Synopsis
- Monoclonal antibody-drug conjugates (ADCs) are a promising therapeutic class, but there are limited methods for their purification, particularly for achieving a specific drug-to-antibody ratio.
- Researchers have successfully applied countercurrent separation (CCS) for the first time to purify an ADC mimic using an aqueous two-phase system (ATPS) composed of specific components.
- The study revealed different partitioning behaviors in the ATPS for the ADC mimic compared to its monoclonal antibody counterpart, suggesting new possibilities for separating high-molecular-weight biomolecules like ADCs.
Article Abstract
Monoclonal antibody-drug conjugates (ADCs) are an expanding therapeutic class of biomolecules for which relatively few analytical and preparative separation options exist. Purification of ADCs with a specific drug antibody ratio is even more challenging. We report the first application of countercurrent separation (CCS) to this problem. An ADC mimic was successfully chromatographed using an aqueous two-phase system (ATPS) consisting of PEG 1000/sodium citrate pH 7.5/water, 17.75/17.75/64.50 (w/w/w). Notably, different partition coefficients () in this ATPS for the ADC mimic (0.09 < < 0.16) and its monoclonal antibody backbone, IgG (0.16 < < 0.27), were observed using CCS. Differential elution behavior of such high-molecular-weight biomolecules, 146,441 vs. ∼150,000 Da, using CCS has no precedent. The results provide a proof of concept for further exploration of the application of ATPSs and CCS to the separation of ADCs.
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| http://dx.doi.org/10.1021/acs.analchem.2c03919 | DOI Listing |
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