HCP Liquid Chromatography-Mass Spectrometry Method
Host cell proteins (HCP) are a class of proteins related to the production process of biopharmaceuticals, which can affect the safety and efficacy of drugs. HCP poses potential safety risks as non-target components in biological products, potentially triggering unknown immune responses in the body that affect the efficacy of biological products, and may even cause hypersensitivity or other adverse reactions. Therefore, it is particularly important to monitor whether the HCP content is within a safe range through effective technical means to improve the safety of biological products.
With continuous technological improvements and upgrades, liquid chromatography-mass spectrometry (LC-MS) is increasingly used for qualitative or quantitative analysis of HCP during purification processes. However, due to the concentration difference between HCP and therapeutic proteins in the solution, which may exceed five orders of magnitude, direct detection using the LC-MS method may be challenging. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) can simultaneously identify and quantify HCP impurities and is the main orthogonal complementary analysis method to enzyme-linked immunosorbent assay (ELISA). By achieving additional separation through high-pH reverse-phase ion exchange or size exclusion prior to LC-MS/MS identification, the LC-MS/MS system can reduce interference from dominant peptides digested from recombinant proteins and maximize the detection of low-abundance HCP-related peptides. LC-MS/MS can provide additional evidence about the HCP clearance process, such as identifying which HCPs are present in the sample or which HCPs have immune reactions with antibodies used for mass spectrometry detection. Moreover, it can reveal the composition and abundance of HCPs at various stages of production, and these methods can also characterize HCPs co-purified with recombinant proteins that do not elicit immune responses in animals.
ELISA is a common technique for analyzing HCP, capable of detecting ppm levels of HCP impurities in milligram-level products, but it cannot provide information about the identity and quantity of individual HCPs in samples. Therefore, LC-MS/MS is needed for mutual validation, complementing the ELISA technique.
BioTech Pack provides biopharmaceutical analysis services, including host cell protein residue ( HCP ) analysis services. BioTech Pack has experienced technicians who can provide a full range of professional services from experimental design, sample testing, to data analysis, and can accurately detect potential residual host proteins in biopharmaceuticals.
Related services:
Antibody-drug conjugates (ADCs) analysis
Protein purity analysis (size exclusion/reverse-phase chromatography)
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