BioResolve™ RP mAb Polyphenyl Columns
BioResolve™ RP mAb Polyphenyl Columns
Waters Enables High-Throughput, Efficient, and Reliable Characterization of mAbs and ADCs
Jacquelynn Smith , Matthew A. Lauber, Ph.D.
Monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) are the fastest growing class of therapeutic agents. There are nearly 50 mAb-related products approved by the U.S. FDA and the European Medicines Agency (EMA) for the treatment of cancer and cardiovascular, neurological, autoimmune, and inflammatory diseases.
Efficient and reliable detection and characterization of microheterogeneities in therapeutic mAbs and ADCs is of critical importance, as sources of variability inherent to the development and manufacturing process have the potential to affect the quality of the final product.1 For Pfizer, improving efficiency and reliability in the process of characterizing protein therapeutics is of critical importance. To help the company achieve its required benchmarks, scientists from Waters took on the challenge of engineering a novel reversed-phase liquid chromatography (RPLC) column for high-throughput separations with enhanced resolution, selectivity, and recovery of target mAbs and ADCs.
Quality Is in the Details
Unlike small molecule drugs, therapeutic mAbs are large, complex biomolecules, typically produced in mammalian cells through recombinant DNA technology. Several factors, including differences between cell lines, or between different clones derived from the same cell line, and variability in post-translational modifications during expression can result in batch-to-batch variability in the final product (Figure 1).2 Some degree of microheterogeneity can be tolerated by patients and is inherent to the process of mAb development, similar to the natural variability occurring in all biomolecules. However, to ensure consistency in the quality of the final product, a set of predefined critical quality attributes that have the potential to influence the quality, safety, and efficacy of a given biologic have to be monitored closely and kept within a certain limit, range, or distribution.
Figure 1. A monoclonal antibody (mAb) and some example variants that are of consequence to structure and function. The BioResolveTM RP mAb Polyphenyl Column is purposefully designed to yield improved separations for deamidation, oxidation, and drug-conjugate variants.
BioResolve™ RP mAb Polyphenyl Columns were designed specifically to address a multitude of technological challenges that are encountered during the characterization of intact mAbs or their subunits by RPLC, including the need for more efficient, higher resolution separations.Scientists from Waters worked to identify the parameters that would lead to enhanced resolution and selectivity, with limited degradation and carry-over, thereby allowing for higher quality, reproducible data. By combining the kinetic benefits of solid-core particle technology with a novel, high-coverage polyphenyl surface chemistry, BioResolve RP mAb Polyphenyl Columns allow users to perform higher throughput, reproducible separations, with enhanced resolution and selectivity for both RPLC and MS-compatible RPLC separations, at lower temperatures and with less acidic ion pairing to limit the degradation of target mAbs (Figures 2 & 3)
Figure 2. A schematic of the BioResolve RP mAb Polyphenyl stationary phase and examples of its benefits.
Figure 3. RPLC chromatograms obtained during the analysis of a mAb subunit sample, wherein the BioResolve RP mAb Polyphenyl Column was used along with lowered temperature and a reduced concentration of acidic ion pairing additive (TFA, trifluoroacetic acid) to minimize on-column degradation and to thereby produce a higher fidelity and more accurate profile of the sample’s composition (purple trace).
