Davy Guillarme previews his presentation at HPLC 2019 highlighting the new trends in LC×LC applied for biopharmaceutical characterization, including the hyphenation with high‑resolution mass spectrometry (HRMS) and ion mobility spectrometry (IMS).
At the previous HPLC 2018 symposium, there was a strong focus on the analytical characterization of protein biopharmaceuticals, including monoclonal antibodies (mAbs) and antibody–drug conjugates (ADCs). In particular, there was an opening plenary lecture from Stacey Ma (Genentech, San Francisco, California, USA) about the most recent analytical technologies used in the biopharmaceutical industry. Additionally, there were also several dedicated lectures and posters sessions on the analysis of mAbs, ADCs, and other formats of proteinâbased drugs.
Since the development of recombinant insulin in 1982, the number of proteinâbased drugs has dramatically increased, and protein biopharmaceuticals have emerged as important therapeutic options for the treatment of cancer, inflammatory and autoimmune disorders, Alzheimer’s disease, and even migraines. Based on this broad range of applications, numerous pharmaceutical companies are increasing their efforts aimed at the research and product development of biologics. Given their obvious benefits in terms of safety and efficacy, mAbs are today the most successful biopharmaceutical products and have totally reshaped the pharmaceutical market (1). As well as mAbs, there is also strong interest in mAb-related products, such as new ADC formats, fusion proteins, PEGylated proteins, bispecific antibodies (BsAbs), antibody fragments (nanobodies and Fab), and polyclonal antibodies (pAb). Last, but not least, some of the first-generation mAbs and fusion proteins will come off patent soon, and there is consequently a significant increase of biosimilars activity requiring powerful analytical methods for comparability assessment of originators and biosimilars.
It is indeed important to keep in mind that these products are highly complex in terms of structure, and the characterization of therapeutic mAbs and ADCs is a tremendous challenge to stateâofâthe-art analytical technologies. Indeed, subtle changes in these large (greater than 150 kDa) molecules can have profound effects on efficacy, pharmacokinetic properties, and toxicity. It is therefore important to have the ability to rapidly and accurately assess changes in the distribution of different isoforms, for example, glycosylation, oxidation, deamidation, lysine truncation of these biomolecules (2).
Today, the most widely used analytical approaches for therapeutic protein characterization are liquid chromatography (LC) and mass spectrometry (MS), probably as a result of the remarkable developments of these strategies in the past few years, which have enabled a new level of performance. Lectures and posters at HPLC 2019 will demonstrate the usefulness of modern LC and MS approaches to demonstrate the product quality and increase the safety of biopharmaceutical products.
As well as the conventional LC modes, such as size-exclusion chromatography (SEC), ion exchange chromatography (IEC), hydrophobic interaction chromatography (HIC), reversedâphase LC, and hydrophilic interaction liquid chromatography (HILIC), it is also possible to combine the most promising chromatographic strategies in a comprehensive two-dimensional liquid chromatography (LC×LC) setup. This allows for a more detailed characterization of some highly complex and heterogeneous new biological entities, thanks to a significantly better peak capacity. In addition, LC×LC also offers the opportunity to combine nondenaturing chromatographic modes requiring significant amount of nonvolatile salts (IEC and SEC) to MS, using a desalting step in the second dimension. Even though LC×LC is becoming more and more popular for the analysis of small molecules, it has only very recently been used for biopharmaceutical characterization (3,4).
During my presentation at the next HPLC 2019 symposium, I will try to highlight the new trends in LC×LC applied for biopharmaceutical characterization, including the hyphenation with highâresolution mass spectrometry (HRMS) and ion mobility spectrometry (IMS).
References
Davy Guillarme holds a Ph.D. degree in analytical chemistry from the University of Lyon, France. He is now senior lecturer at the University of Geneva in Switzerland. He has authored more than 200 journal articles related to pharmaceutical analysis. His expertise includes HPLC, UHPLC, HILIC, LC−MS, SFC, and the analysis of proteins and mAbs. He is an associate editor of Journal of Chromatography B and editorial advisory board member of several journals.
2024 EAS Awardees Showcase Innovative Research in Analytical Science
November 20th 2024Scientists from the Massachusetts Institute of Technology, the University of Washington, and other leading institutions took the stage at the Eastern Analytical Symposium to accept awards and share insights into their research.
Inside the Laboratory: The Richardson Group at the University of South Carolina
November 20th 2024In this edition of “Inside the Laboratory,” Susan Richardson of the University of South Carolina discusses her laboratory’s work with using electron ionization and chemical ionization with gas chromatography–mass spectrometry (GC–MS) to detect DBPs in complex environmental matrices, and how her work advances environmental analysis.
AI and GenAI Applications to Help Optimize Purification and Yield of Antibodies From Plasma
October 31st 2024Deriving antibodies from plasma products involves several steps, typically starting from the collection of plasma and ending with the purification of the desired antibodies. These are: plasma collection; plasma pooling; fractionation; antibody purification; concentration and formulation; quality control; and packaging and storage. This process results in a purified antibody product that can be used for therapeutic purposes, diagnostic tests, or research. Each step is critical to ensure the safety, efficacy, and quality of the final product. Applications of AI/GenAI in many of these steps can significantly help in the optimization of purification and yield of the desired antibodies. Some specific use-cases are: selecting and optimizing plasma units for optimized plasma pooling; GenAI solution for enterprise search on internal knowledge portal; analysing and optimizing production batch profitability, inventory, yields; monitoring production batch key performance indicators for outlier identification; monitoring production equipment to predict maintenance events; and reducing quality control laboratory testing turnaround time.
Infographic: Be confidently audit ready, at any time and reduce failures in pharma QC testing
November 20th 2024Discover how you can simplify the audit preparation process with data integrity dashboards that provide transparency to key actions, and seamlessly track long-term trends and patterns, helping to prevent system suitability failures before they occur with waters_connect Data Intelligence software.
Critical Role of Oligonucleotides in Drug Development Highlighted at EAS Session
November 19th 2024A Monday session at the Eastern Analytical Symposium, sponsored by the Chinese American Chromatography Association, explored key challenges and solutions for achieving more sensitive oligonucleotide analysis.