The Column June 2017

June 2017 | Volume 13, Issue 9
Digital Edition
By LCGC Editors
This issue of The Column in its original digital magazine format.
Cover Story
A novel quantitative method combining multiple reaction monitoring (MRM) and ultrahigh‑pressure liquid chromatography coupled to mass spectrometry (UHPLC–MS) has been developed by Carlito Lebrilla and his team to study site-specific glycosylation in manufactured recombinant monoclonal antibody drugs (rmAbs). He spoke to The Column about this research.
Features
As a result of the pharmaceutical cGMP for the 21st century and quality by design (QbD) initiatives championed by regulators, the biopharmaceutical industry has been looking for ways to introduce more automated and higher information content analyses into manufacturing, late-development, and quality control (QC). Mass spectrometry (MS-) based attribute monitoring assays have been proposed as key tools to provide the sensitivity, throughput, selectivity, and flexibility required for monitoring critical product and process attributes for biopharmaceutical production and release. Two analytical workflows, subunit multi-attribute monitoring (MAM) and peptide MAM, have emerged to dominate this discussion, and this article is intended to reflect on the active debates over the needs, challenges, and practical limitations for adopting MS-based attribute monitoring for late-development and QC.
An increasing number of drugs coming onto the market are proteins rather than small molecules. A major portion of these are produced using a host cell system. Host cells express many of their own proteins that can easily contaminate the recombinant protein drug. Traditionally, these host cell proteins (HCPs) have been measured using immunoassays, but recently, orthogonal analytical methods, particularly mass spectrometry (MS), have started to be used. This article considers some of the current methods for HCP detection, with a focus on MS.
Antibody–drug conjugates (ADC) are an emerging pharmaceutical technology, with the potential of being the true “magic bullet”. However, these inherently complex biomolecules have unique analytical challenges. One is the determination of the drug-to-antibody ratio (DAR). The average DAR and the drug distribution need to be monitored and could determine the success of an ADC. This article discusses the chromatographic methods used to determine DAR, including hydrophobic interaction chromatography (HIC), reversed phase, and liquid chromatography–mass spectrometry (LC–MS).
Dynamic light scattering (DLS), widely recognized for its ability to detect aggregation in purified protein solutions, is actually far more versatile and may be applied to the evaluation of stability and viscosity of therapeutic proteins and other biopharmaceuticals. In a fully automated, microwell plate reader format, DLS constitutes a high-throughput biophysical screening technique for early-stage assessment of candidate developability and optimal formulation conditions.
Biotherapeutic proteins, such as monoclonal antibodies (mAbs), are heterogeneous and exist as variant mixtures of structurally similar molecules. The heterogeneity of monoclonal antibodies is revealed by charge-sensitive methods, such as ion exchange chromatography (IEX). Changes in charge profile can significantly impact the structure, stability, binding affinity, and efficacy of the biotherapeutic drug. It is therefore necessary to understand the profile of the drug so that variants are identified and controlled. This article describes advances in ion exchange column chemistries, elution buffers, and ultrahigh-pressure liquid chromatography (UHPLC) instruments to meet the needs for modern, robust analysis of charge variants in monoclonal antibodies and therapeutic proteins.
News
Thermo Fisher Scientific has been selected as a partner of INTELLItrace Work Package (WP) 18, part of the European Food Integrity Project aimed at developing a validation of untargeted methods to assure the quality, authenticity, and safety of the food chain.
Researchers have developed and validated a stability- and potency-indicating assay protocol for high-throughput quality assessment of vaccines consisting of recombinant virus- “like” particles (VLPs) using conformation-dependent antibodies coupled to size-exclusion high performance liquid chromatography (SE-HPLC).
Fluence Analytics, formerly Advanced Polymer Monitoring Technologues (APMT), has announced an increase in investment led by Energy Innovation Capital (EIC) but also including participation from other investors.
Researchers from the Chinese Academy of Science in Beijing, China, have used high‑performance size-exclusion chromatography (HPSEC) and differential scanning calorimetry (DSC) to study the stabilization of inactivated foot and mouth disease virus.
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