Monoclonal Antibodies: Analysis & Characterization
Monoclonal antibodies (mAbs) have emerged as revolutionary therapeutic agents for an array of human diseases. In addition to their large mass and complex structure, mAbs are varied in their origin, makeup, effector function and delivery, and therefore require thorough formulation development, including characterization, quantitation and preservation. Quality control is critical to maintaining not only the product's quality, but also safety–mAbs are sensitive to the manufacturing process and precision during manufacturing is essential to avoid variations in the product protein.
Regulatory agencies require that all biopharmaceutical companies monitor critical quality attributes (CQA) for the commercialization of any mAb therapeutic product. Accurately doing so is dependent on the sensitivity and resolution of your analytical system. During mAb development, the analysis should afford high sensitivity to post-translational modifications, such as glycosylation and phosphorylation, and should adequately resolve the size/fragment heterogeneity often inherent in complex protein therapeutics.
Charge Variant Analysis of Monoclonal Antibodies
Imaged capillary isoelectric focusing (icIEF) technology is widely regarded as the standard methodology for separation of proteins or peptides based on their isoelectric points (pIs). During icIEF, a capillary is filled with the study sample and an ampholyte mixture. Upon the application of an electric field, the ampholytes form a pH gradient in the capillary. The charged protein migrates through the pH gradient in response to the applied voltage until it reaches the pH value that is equivalent to the protein's pI, at which point the migration stops due to a net neutral charge. In this way, each protein in a sample becomes "focused" per its specific pI. The entire capillary is then imaged to generate a charge heterogeneity profile of the sample under study. pI values and relative peak areas are examined to create sample profiles. icIEF profiling can be applied throughout the manufacturing process of mAbs from scale-up to formulation development and stability testing, and quality assurance (QA) of lot-to-lot consistency, ensuring the necessary quality attributes of the mAb.
The Maurice™ system brings icIEF technology to the next level, by vastly simplifying your charge heterogeneity analysis workflow. There's no transfer line, capillary and switch valve maintenance, or tedious cartridge installation procedures at the beginning of your run. Just load your cartridge, samples and reagents into Maurice, set up your batch parameters, hit Start and you're done. In addition to UV detection applied in earlier icIEF systems, Maurice's native fluorescence detection for cIEF measures the fluorescence emission of tryptophan's aromatic group for improved signal/noise ratio. It's label-free so you're not wasting time optimizing protein labeling or dealing with the background noise that results when a label unconjugates from your protein. Baselines are significantly cleaner and less sensitive to ampholyte interference, giving you more options when optimizing your pH gradient.
Download our application note to learn how Maurice improves charge variant analysis with his native fluorescence.
Characterization of Monoclonal Antibody Size Variants
mAb fragmentation and size migration assessments have evolved from gel (SDS-PAGE) to capillary electrophoresis (CE-SDS). CE-SDS is a high-throughput analytical technology equipped with quantitative data integration that has effectively reduced turnaround time during screening and development. CE-SDS separation and analysis with Maurice lets you assess the purity and identity of therapeutic monoclonal antibodies using a much simpler workflow that results in highly robust, high-quality data.
Download our application note to learn how Maurice's simple workflow lets you monitor the degree of heavy chain glycosylation and degree of fragmentation in reduced and non-reduced IgG molecules quickly and easily.
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