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Host Cell Protein Analysis
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Host Cell Protein Identification (HCP)
Host cell proteins (HCPs) are impurities in drug products regularly created during the manufacturing process. HCPs are typically low-level endogenous proteins derived from the host organism which can often be difficult to purify or detect, yet can affect therapeutic activity and stability of the drug. HCP characterization is an essential process in biopharmaceutical manufacturing to ensure the safety, efficacy, and quality of therapeutic products. Mass spectrometry (MS) has become a powerful technique used for this purpose due to its sensitivity and accuracy.
CovalX_Host-Cell-Protein In recent years, mass spectrometry has become a reference technique for HCP analysis. High resolution and advanced data analysis softwares can provide a more accurate characterization of the HCPs without affinity-based bias as with traditional antibody-based techniques such as ELISA and Western blot.
Using the latest high-resolution nano-liquid chromatography electrospray (nLC-ESI) mass spectrometry and data analysis softwares, CovalX can detect and characterize these impurities with the highest degree of certainty.
DALL·E 2024-09-06 09.52.30 - An image of CHO cells (Chinese Hamster Ovary cells) viewed under a microscope, depicted in deeper shades of blue. The cells should appear rounded with What are Host Cell Proteins?
Host Cell Proteins (HCPs) are proteins produced by the host cells used in the production of biopharmaceuticals. For instance, recombinant proteins are often expressed in mammalian cells, yeast, or bacteria. During the production process, the host cells produce their own proteins, which can be co-purified with the target therapeutic protein.
Mass Spectrometry for HCP Characterization
1. Sample Preparation
- Protein Digestion: Special proteolysis protocol is used to generate peptides from HCP
- Purification: A purification protocol separates the undigested mAbs from HCP peptides.
2. Mass Spectrometry Analysis
- Peptide Separation: HCP peptides are separated using nano flow Liquid Chromatography (nLC) mass spectrometry.
- High-Resolution Mass Spectrometry: HCP peptides are ionized and detected providing an accurate mass measurement.
- Tandem MS (MS/MS): Multiple stages of mass spec fragmentation generate the most accurate peptide and protein identification.
3. Data Analysis
- Database Search: The acquired mass spectra are compared against protein databases to identify the HCPs present.
- Quantification: The relative abundance of each HCP can be determined by comparing the intensity of peptide signals in the mass spectra.
- Bioinformatics Tools: Specialized softwares are used to interpret the complex data, helping to identify HCPs and quantify their levels.
Why Characterize HCPs?
- Regulatory Compliance: Regulatory agencies like the FDA and EMA require rigorous testing to ensure that HCPs are controlled in therapeutic products.
- Product Efficacy: High levels of HCPs can affect the efficacy of the therapeutic protein, potentially leading to issues with affinity, stability and immunogenicity.
- Process Optimization: Understanding HCP profiles helps in refining production processes to ensure quality manufacturing.