Host Cell Protein (HCP) Analysis

We offer comprehensive HCP analysis to detect and quantify HCPs in the high complexity samples containing both HCPs and Drug Substance (DS). Our HCP Assay by 2D DIGE, 2D WesternBlot and HCP Assays by Nano LC-MS/MS platforms include HCP Antibody Coverage, HCP profiling and Product purity. These HCP analyses allow high sensitivity detection and accurate quantitation of trace amounts of HCP contaminants in biopharmaceutical proteins.

HCP Assays by 2D DIGE

PlatformSeparate byDetectionSensitivityProtein imageProtein/WB
in 1 gel		Advantages
1D DIGEMWCyDye label0.2 ngImageImage1) Quick comparison of
HCPs in one dimension
1D DIGE WBMWCyDye label0.1 ngImageImage
2D GelMW and pIStaining1-100 ngImageImage1) Detailed comparison of
HCPs in two dimensions
2) Accurate quantitation
2D WBMW and pIStaining0.1 ngImageImage
2D DIGEMW and pICyDye label0.2 ngImageImage1) Detailed comparison of
HCPs in two dimensions
2) Accurate quantitation
3) High sensitivity
4) High consistency
5) High dynamic range
2D DIGE WBMW and pICyDye label0.1 ngImageImage

HCP Assays by Nano LC-MS/MS

PlatformSeparateDetectionSensitivityProtein imageProtein/WB
in 1 gel		Advantages
NanoLC
MS/MS		Hydrophobicity
& MW-optional		MS/MS10 amolImage1) Quick comparison of
HCPs in one dimension

FAQ

Frequently asked questions about HCP advantages, process & time:

Our 2D Western platform, employing fluorescent labeling, can generate both HCP and WB images from the same gel and same membrane. Our report provides HCP, Western blot and HCP/Western overlay images, allowing the direct visualization of comprehensive antibody coverage that ELISA or LCMSMS cannot provide. In addition, our large format 2D offers high resolution of HCPs and their modified forms that ELISA or LCMSMS cannot achieve, greatly improves the sensitivity and accuracy of coverage.

AAE, based on antibody immuno-binding with antigen under the native condition, has limitations in quantitating antibody coverage:

  • False positive: a) Proteins directly and indirectly associated with antibody-bound proteins; b) antigens present in the HCP antibody; c) co-purified serum proteins in HCP antibody
  • False negative: a) Protein complex may block antibody binding; b) some bound HCPs or their degraded fragments cannot be eluted
  • In addition, AAE still needs be coupled with downstream LCMSMS or 2D Western blot, such lengthy process introduces more variations and inconsistency.

7-9 days

No

3 samples in 1 test. Our report provides individual HCP image and overlay image of different HCP samples, giving you the direct visualization and comparison of HCP compositions, modifications and fragmentations that no other platform can achieve.  In addition, the quantitation analysis provides spot counting, similarity analysis, total HCP and DS quantity. 

Yes. Based on the molecular weight and pI of the DS, we can modify IEF and 2D conditions to achieve the optimal separation of HCP from DS.