host cell protein analysis mass spectrometry

1. Host cell protein analysis mass spectrometry Host that isoelectric point, hydrophobic properties, and structures. Figure 1 shows a harvested product pool of bacterial and mammalian fermentation by two-dimensional difference gel electrophoresis (2D-DIGE). Identification and quantification HCPs is very important in the development of biopharmaceutical products to ensure product purity and patient safety. Protease of HCPs can influence the protein pattern in culture and influence the purification In addition, HCPs can undergo post-translational modifications that quantification and characterization more difficult. Moreover, high HCP variance results in different biochemical properties which could cause an immune response or allergic reactions even anaphylactic shock. Cell Proteins (HCPs) are a heterogeneous, complex group of proteins derived from the host cell. HCPs differ significantly in molecular mass, process. make 2D-DIGE silver stain of a harvested product stain of a harvested product Figure 1 2D-DIGE silver Although contaminate such as, species, there appropriate methods other impurities for biopharmaceuticals. examine each protein drug on a case-by-case maximum acceptable levels of HCPs and patient the host cells contain thousands of HCPs and that could potentially the final product and the host cells can be various organisms, bacteria, yeast and cell lines derived from mammalian or insect is no explicitly guideline that describes the selection of for characterization and quantification of HCPs and Moreover, the regulatory agencies basis to determine appropriate risk. The quantification manufacturing. The either immune-specific non-specific methods, such as, qualitative or semi-quantitative method purification and for lot release. However, and characterization, the ideal method not recognize all HCPs species, but also should time-saving and labor-saving. selection of appropriate test methods for identification and/or of HCP have been a concern during process development and methods used for HCP analysis can be classified as methods, such as, western blotting and ELISA, or electrophoresis and MS. In general, a would be sufficient for sample during process of drug development only should have the capability to be quantitative, high-throughput, A presented range of methods for the detection and characterization of HCPs are in Creative Proteomics (Table 1). Among these, ELISA,

2. electrophoresis, and MS are sometimes they are used in combination for the most common techniques and are detailed characterization. Table 1, Overview methods of HCPs methods Overview methods of HCPs methods Technology platform 1D/2D - Silver stain/Coomassie blue/SYPRO Ruby/CyDyes/Autoradiography 1D/2D (Non-target,shotgun) - Silver stain/Coomassie blue - HPLC - Mass spectrometry 2D HPLC-mass spectrometry (Non-targeted, shotgun) HPLC-mass spectrometry (target MRM) Summary Identification / quantification of HCPs Report A including the sample preparation, experimental procedure and instrument parameters. detailed technical report will be provided at the end of the whole project, The reported in the report. stain results, protein list (FDR<1%)/and concentration/amount will be Ordering Procedure: Ordering Procedure Our customer service representatives are available 24 hours a day, from Monday to Sunday