Critical Quality Attributes (Primer)

The future of biopharmaceuticals looks promising with life changing treatments. The field keeps growing and is powered by innovative, ground-breaking therapies in cancer treatment and auto immune diseases. Advancing these novel biotherapeutics safely in the clinic requires reliable manufacturing and quality control processes. The complex heterogenous nature of biotherapeutics requires accurate and robust analytical testing methodologies with dependable chromatographic separations. Identifying critical quality attributes (CQA) is the most difficult step in implementation of quality by design (QbD) for development and production of biopharmaceuticals. Defining each product attribute is extremely challenging and therefore, consistency of product quality becomes even more important. We at Agilent CrossLab, designed and manufactured our AdvanceBio columns and consumables to match our customers’ needs. In this compendium, we have therefore selected applications to illustrate the state of-the-art chromatographic separation for each CQA using either HPLC-UV or light scattering detectors. We provide an overview of different chromatographic separation technologies for each CQA using the diverse range of chemistries with in our biocolumns portfolio. There are examples of reversed-phase, size exclusion, ion exchange, and hydrophilic interaction chromatographic analyses of therapeutic proteins, mAbs, and antibody-drug conjugates. Let us help you improve your productivity, method robustness, and reliability of your analytical results. We want to stand by you on this journey in developing safer and effective biotherapeutics. Your success is our success Biotherapeutic proteins are highly complex molecules, which are typically produced by fermentation using recombinant methodologies. This production process however, results in the generation of many different variants of these proteins. Ensuring the quality of such materials is paramount. This means confirming the product is correctly manufactured, any impurities are identified and quantified, and the potency of the protein is determined. As a result, it is necessary to perform tests on the intact, nondenatured molecule. Something as large as a monoclonal antibody may contain more than 1,300 individual amino acids and have a mass of more than 145,000 daltons. However, identifying a single minor impurity such as deamidation of asparagine resulting in a mass difference of just one dalton, which may occur at any of perhaps twenty or more different asparagine positions throughout the molecule, is challenging. Only by breaking down the molecule into fragments (such as light and heavy chains) and then into smaller polypeptide chains through enzymatic treatment is it possible to begin to pinpoint some of these subtle differences. Many different types of variant can be created and these are often referred to as posttranslational modifications, or PTMs. They arise after the protein has been expressed, and can be a consequence of the manufacturing conditions, or exposure to conditions that cause changes to occur. Fluctuations in temperature, pH, concentration, or exposure to enzymes can all lead to variants developing. Glycosylation in particular is highly variable but is of major importance to the efficacy of many proteins. Understanding the different types of impurity and the risks each pose forms the basis of Critical Quality Attribute (CQA) monitoring The purpose of this document is to highlight some of the HPLC applications suitable for the different aspects of CQA monitoring. As well as providing guidance for the appropriate liquid chromatography column for the different types of detection that may be required, and to provide a valuable reference for future consideration. Affinity chromatography Titer Determination Ideal for mAb titer determination during process development Glycan Analysis Hydrophilic interaction chromatography H2O H2O H2O H2O Fast, high-resolution, reproducible glycan separation Charge Variant Analysis Ion exchange chromatography Enhances the accuracy and speed of biomolecule characterization CO2 – CO2 – CO2 – Amino Acid and Cell Culture Analysis Small molecule chromatography (<150 Å) H H H H R O O N C C Delivers robust, high-resolution separations Intact and Subunit Purity Large molecule chromatography (>150 Å) Selectivity options for every separation need Aggregate/ Fragment Analysis Size exclusion chromatography Accurate, precise quantitation for a broad range of biomolecule separations Peptide Mapping Za odpiranje dokumenta
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