Delaney Doran, Matthew Myers, Katarina Moravcevic, Steven LaBrenz, Pinger Wang, Xiaoying Ji
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The Hidden Influence: Impacts of Residual Dimethylformamide in NDSB-211 on icIEF Separation for Monoclonal Antibodies.
Monoclonal antibodies (mAbs) present analytical challenges due to their inherent heterogeneity and susceptibility to post-translational modifications (PTMs) during production and storage. Monitoring of charge heterogeneity profiles by imaged capillary isoelectric focusing (icIEF) has been aided by the use of non-detergent sulfobetaines (NDSBs), particularly NDSB-211, to enhance protein solubility and stability. When used in a quality control laboratory setting, NDSB-211 has shown performance variability over time due to residual manufacturing impurities that impact the capillary isoelectric focusing separation. Dimethylformamide (DMF) was detected using 1H nuclear magnetic resonance (NMR) in several lots of NDSB-211 that simultaneously produced poor resolution of basic variants of a mAb on icIEF. A parallel spiking study using DMF at 1%-10% (v/v) on 1H NMR and icIEF illustrates the interference in charge heterogeneity separation when a large amount of DMF is present in the NDSB-211 reagent. As DMF is not found on the NDSB-211 certificate of analysis from any vendor, more traditional additives for icIEF are suggested to limit the potential interference for analytical methods.
期刊介绍：
ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.).
Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences.
Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases.
Papers describing the application of standard electrophoretic methods will not be considered.
Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics:
• Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry
• Single cell and subcellular analysis
• Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS)
• Nanoscale/nanopore DNA sequencing (next generation sequencing)
• Micro- and nanoscale sample preparation
• Nanoparticles and cells analyses by dielectrophoresis
• Separation-based analysis using nanoparticles, nanotubes and nanowires.
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