CE Pharm 2021

Sponsored by ProteinSimple, a Bio-Techne Brand

The first segment of the technical seminar will focus on an innovative fractionation workflow. A thorough characterization of a biopharmaceutical drug candidate is critical during development to attain the ultimate goals of commercializing a safe and efficacious therapeutic for patients. An important regulatory requirement is to characterize charge variants. The traditional technique used by Maurice icIEF users is to develop an ion exchange chromatography assay, collect fractions and analyze them using a mass spectrometer. ProteinSimple has developed a fractionation capable icIEF solution and is collaborating with 908 Devices in charge variant characterization using their ZipChip device connected to a mass spectrometer, providing a complete workflow for charge variant analysis. This solution offers a powerful charge variant characterization tool for biotherapeutic characterization. The presentation will include the analysis of the NIST mAb along with a second monoclonal mAb and will present icIEF fractionation and characterization data.
The second segment will discuss a unique fast and high throughput CE-SDS Maurice capability. Scientists currently use techniques that can require up to 45 minutes per injection to be able to analyze a sample and achieve excellent resolution. The innovative Turbo CE-SDS™ Cartridge for Maurice provides results in less than 6 minutes (up to a 75% time reduction from some traditional assays) and provides similar profiles and % area values compared to traditional methods. Reproducibility is excellent with CVs as low as 0.05%. This new cartridge, which runs on Maurice after a simple software upgrade, enables analysis of up to 144 samples in 8 hours across 2 batches, with up to 96 injections per batch. This high throughput CE-SDS cartridge will enable the screening of a large number of samples on CE-SDS methods addressing the sample bottle-neck challenge that customers are currently facing. The speed and data quality also make it ideal for running a much smaller number of samples.
Attendees will learn about:

o The new charge variant characterization solution, which offers workflow enhancements
o How the new Turbo CE-SDS™ cartridge for Maurice provides a fast and high throughput capability

Sponsored by 908 Devices

Characterization of biologics, especially mAbs is a challenge task in order to ensure safety, efficacy and potency of a therapeutic agent due to the heterogeneity of the structure happened throughout the steps of cell culture process, purification process and storage. The presence of positively charged and negatively charged amino acids and negatively charged glycans (sialic acids) means that mAbs exist as multiple charged species. Understanding which amino acids or glycans are involved and their specific location within mAb is of paramount importance. Traditional ion exchange chromatography can enable the separation of some charge variants, particularly those positioned on the surface of the protein. However, microfluidic capillary electrophoresis-mass spectrometry (microfluidic CE-MS) to analyze intact mAb and to assess the root cause of the stressed mAb provides a very powerful direct and generic tool for separation and identification of charge heterogeneity of mAbs.

Sponsored by Intabio, Inc.

Biotherapeutics are a complex and heterogeneous class of drug molecules that require comprehensive and rapid characterization throughout the development process. Charge heterogeneity of monoclonal antibodies (mAbs) can result from cellular processes, chemical degradation and production conditions during manufacturing, which may impact the efficacy, safety and potency of therapeutic mAbs. Imaged capillary isoelectric focusing (iCIEF) is an established platform technique for separating and quantifying charge variants based on changes to the molecule’s isoelectric point that result from post translational modifications (PTMs). However, identifying those PTMs that result in changes to the iCIEF profile often requires fractionation, LC method development, offline MS characterization and reanalysis by iCIEF, which is a lengthy and laborious process.

To overcome these bottlenecks, we describe advances in microfluidic chip technology development that have enabled the direct coupling of iCIEF with MS to provide charge variant separation, UV quantitation and MS characterization. These advancements provide unprecedented insights into critical PTMs, including glycan analysis, charge variant peak ID and intact mass analysis of biotherapeutic mAbs. Several examples of charge variant characterization demonstrating high-resolution iCIEF separation coupled with sensitive detection and mass identification of major and minor proteoforms will be highlighted.

This technology holds promise to reduce the timeline, investment and risk associated with the development of biotherapeutics.