Within a rapidly growing biopharmaceutical market, there is an increasing demand for the development of a rising number of biotherapeutic protein drugs, such as monoclonal antibodies (mAbs). During the development and production of biotherapeutic proteins, typically using mammalian cell lines, such as Chinese hamster ovary (CHO) cells, additional proteins are expressed as well by the host cell. These so-called host cell proteins (HCPs) will need to be removed, as they can influence the safety and efficacy of the biotherapeutic drug. However, a variety of these HCPs can be co-purified with the target protein, and end up in the biotherapeutic end-product.
Regulating agencies will therefore require the monitoring of HCP levels in the final product. After multiple purification steps, the HCPs presence in the end product will typically be in low concentrations. Enzyme-linked immunosorbent assays, or ELISA, has been the technique of choice for HCP analysis, but today liquid chromatography coupled to mass spectrometry is more and more used, especially as a complementary technique to ELISA.
Looking at the complexity of HCP mixtures, micro Pillar Array Column (µPAC™) technology can play a key role in the LC-MS analysis. Taking full advantage of the highly ordered pillar structure that forms the column backbone, highly sensitive and reproducible analysis of such low concentration HCPs can be routinely performed. In combination with high resolution accurate mass measurement mass spectrometry, such as the Orbitrap Q-Exactive HF-X, a sensitive and selective HCP monitoring workflow can be designed.
- Identify and quantify Host Cell Proteins by LCMS
- Selective and sensitive LCMS characterization of low concentration HCPs
- Get to know your antibody process impurities