May 16th 2024
Scientists from the Lunan Pharmaceutical Group explored the use of UHPLC-MS/MS to test the effects of Shouhui Tongbian capsules (SHTBs) on slow transit constipation (STC).
Intact Mass Analysis of Biopharmaceuticals as its Own Unique Application
November 1st 2021Intact mass analysis is becoming increasingly useful for characterizing biologics. We describe the current application of intact mass analysis, including quantitation, sequencing, and structural characterization.
New Aspects in the Integration of Mass Spectrometry Technologies in the Biopharmaceutical Industry
October 1st 2021Recent developments in high-resolution mass spectrometry (HRMS) are aiding biopharmaceutical development and simplifying routine monitoring, with applications in areas like multi-attribute methods, coupling established purity methods with MS, and subunit analysis.
Tools for Functional Assessment of Biotherapeutics
June 1st 2021We present the main analytical techniques for performing functional characterization of biotherapeutic products. Such assessments are particularly critical for biosimilars, where analytical testing must ensure functional comparability with the innovator product.
We explore the impact of two different stationary phases and ion-pair reagents on the retention behavior of a therapeutic peptide using reversed-phase liquid chromatography. This information is of fundamental importance for the development of reliable, selective, and fast analytical methods able to separate and identify the target peptide.
Online Hydrophobic Interaction Chromatography–Mass Spectrometry (HIC–MS) Analysis of Proteins
February 1st 2021Recent advances in stationary phases for hydrophobic interaction chromatography (HIC) permit HIC–MS analysis of intact antibodies and other proteins using direct flow to the mass spectrometer.
The Multi-Attribute Method (MAM) for the Characterization of Biopharmaceuticals
January 1st 2021New analytical workflows are needed to address the advances in biopharmaceutical product composition. A description of the multi-attribute method (MAM) is given, which has been developed to monitor critical quality attributes (CQAs) simultaneously and directly.
In Search of The Needle in The Mariana Trench: Host Cell Proteins and the Problem of Dynamic Range
October 9th 2020This article explores the analytical challenges associated with HCP monitoring and reviews recent advances in HCP characterization, with special emphasis on high performance liquid chromatography mass spectrometry (HPLC–MS)-based methods and HCP enrichment techniques.
Analytical Separation Methods for Therapeutic Oligonucleotides
October 9th 2020The diverse nature of oligonucleotide therapeutics leads to the requirement for multiple separation methods to characterize quality attributes. Highly chemically-modified to improve efficacy and resilience to nucleases, they are challenging to analyse using a single method. This article discusses multiple chromatographic separation methods, and the benefits of mass spectrometry (MS).
Physicochemical Methods for Vectors and Ancillary Materials in Cellular and Gene Therapies
October 9th 2020Cellular and gene therapies (CGTs) have contributed significantly to the improvement of clinical outcomes for patients in the recent years. This paper discusses a range of physicochemical methods that play an important role in the difficult characterization of viral vectors, to meet the unique needs of CGT manufacturing process development, process and product characterization, and the quality control testing of these materials.
Rapid Adeno-Associated Virus Genome Integrity Analysis by Capillary Gel Electrophoresis
October 9th 2020In this paper, two adeno-associated virus (AAV) genome integrity analysis workflows are introduced; a standard sample preparation protocol and an accelerated procedure, both utilizing capillary gel electrophoresis with laser induced fluorescent detection (CGE–LIF) to analyze the released nucleic acids.
Troubleshooting LC Separations of Biomolecules, Part II: Passivation and Mobile-Phase Additives
August 1st 2020If you are analyzing metal-sensitive biomolecules, and a bioinert instrument is unavailable, or insufficient, passivation or mobile-phase additives may help. Here’s how to use those solutions, with tips for avoiding potential pitfalls.
Practical Two-Dimensional Liquid Chromatography in Drug Metabolism Studies and Bioanalysis
June 30th 2020Filip Cuyckens from Janssen R&D in Belgium spoke to LCGC Europe about recent innovative approaches he and his team developed to support drug metabolism and pharmacokinetic studies, and the inventive role that two-dimensional liquid chromatography (2D-LC) plays in his laboratory to boost sensitivity, solve recovery issues, and increase overall efficiency.
Boosting the Purification Process of Biopharmaceuticals by Means of Continuous Chromatography
Single-column (batch) chromatography, involving two or more successive single-column (batch) chromatographic steps, is a standard approach for purifying biopharmaceuticals. Step one, known as the capture step, is used to remove product-related impurities, and step two, the polishing step, is used to remove product-related impurities. Here we present and illustrate the advantages of continuous chromatography for these separations: capture simulated moving bed (captureSMB) for the capture step and multicolumn countercurrent solvent gradient purification (MCSGP) for polishing.
Multidimensional Separation Techniques for Characterization of Biotherapeutics
June 1st 2020Multidimensional separations, in which two or more separation methods are coupled, are a valuable analytical tool for higher peak capacity and improved selectivity for the analysis of complex samples like biotherapeutics.
Glycosylation Analysis Through Released N-Glycan Workflows
May 6th 2020Post-translational modifications are potential critical quality attributes (pCQAs) routinely assessed in biotherapeutic development. Glycosylation is one of the most important attributes to assess because it affects protein function as well as antigen receptor binding. N-glycosylation of asparagine residues is the most common pCQA assessed during monoclonal antibody (mAb) therapeutic development. There are a few protocols to assess and quantitate N-glycans, but the most common approach is through an enzymatic release and labelling procedure, followed by separation and detection. This article demonstrates the method development considerations for sample preparation and chromatographic analysis of N-glycans of therapeutic mAbs.
A method was developed for the molecular weight characterization of heterogeneous polymer mixtures, such as heparins and glatiramer acetate, noting that single molecular structures are not adequate for creating a molecular weight calibration curve. That limitation is overcome in this work, which demonstrates method validation and application to process samples.