HCP profiling for bioprocess optimization

Do you want to improve and speed up your process development?

The best way to identify Host Cell Proteins (HCPs) in your process samples is by LC-MS analysis. For bioprocess optimization, you will benefit from an optimized workflow providing reproducible HCP profiling through identifying and quantifying individual HCPs.

The HCP analysis provides the identity and quantity of each HCP, in addition to the total HCP content (ng HCP/mg drug substance). Thus, the results can be used for data-driven optimization of downstream purification to remove process-related impurities and bring down HCP levels.

HCP profiling for all expressions systems and types of samples

One of the biggest advantages of LC-MS-based HCP analysis is that the same assay can be applied to all expression systems and all types of samples – from early harvest to pure drug substance. Thus, HCPs can be determined in, e.g., the four main cell-based expression systems: bacterial, yeast, insect, and mammalian. The only requirement is that the organism’s proteome needs to be sequenced.

Common yeast hosts include Pichia pastoris, Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Hansanuela polymorpha. Besides Escherichia coli, which we often analyze, common bacterial hosts include Lactococcus lactis, Bacillus subtilis, Pseudomonas fluorescens, Staphylococcus aureus, and Brevibacillus choshinensis. Insect cell lines, such as SF9, SF21, Autographa californica, Lepidoptera, and Spodoptera frugiperda, are also worth mentioning. Finally, we see many cell lines of mammalian origin, including CHO, HEK 293, HeLa, L929, BHK, Y79, U2OS, HepG2, NIH3T3, NS0, P19, CAD, MCF-7, and J558L.

The standard HCP profiling service includes

• Generic sample preparation for process samples in different matrices
• LC-MS peptide analysis on a robust microflow HPLC and Sciex TripleTOF 6600 mass spectrometer using SWATH mass spectrometry (LC-MS/MS) for reproducible HCP identification and quantification
• Report with a list of HCPs and their physiochemical properties (MW, pI, AA sequence, database accession number) for each process-sample – for use in your bioprocess optimization.

Send us a description of your project, and we will be glad to discuss HCP profiling analysis with you.