Host Cell Protein (HCP) Analysis

Host cell proteins (HCPs) are host cell-derived proteins introduced during the manufacturing process of recombinant therapeutic products. These residual HCPs could affect the efficacy, safety and product quality.

Applied Biomics’ HCP analysis complies with U.S. Pharmacopeia general chapter 1132.1 regarding  accuracy, sensitivity, dynamic range and specificity. We offer both Gel-based (1D and 2D) and Mass spectrometry-based (LC-MS/MS) analysis to detect, identify and quantify HCPs for samples at different stages of process. Our services offer orthogonal methods to compliment your in-house tests and facilitate regulatory filing.

To better serve our customers, we now offer Anti-HCP Antibodies (CHO, HEK, E. coli) with over 95% HCP coverage.

Please use the table below to guide you in choosing the right service:

HCP ApplicationsStudy ObjectivesSamples
per Test		Data Report
HCP ImageQuantSensitivityExclude DS
Process Optimization & HCP Characterization 1
1D HCP ProfilingQuick comparison of HCPs by Mw1-10xSemi10-100
2D HCP ProfilingIn-depth comparison of HCPs
by Mw and pI		1-3xx10-100Opt
Evaluate HCP Antibody Coverage 2
2D HCP Antibody CoverageAssess/compare commercial vs. process specific HCP Ab1xx10-100Opt
HCP Impurity Characterization 3
HCP Identification and Quantitation by LCMSMSIdentify and quantify all HCPs in a sample1x1-10
Advanced HCP Analysis
Advanced Analysis OptionsQuadrant Analysis, DS Exclusion, Negative Spots Exclusion1-3xx10-100Opt

1. Process Optimization & HCP Characterization:

For in-depth analysis of HCP clearance during the process development, our 2D HCP profiling can test up to 3 samples in each run and generates comprehensive HCP 2D image and quantitation of each sample as well as comparisons between the samples.  Our 1D HCP profiling offers a rapid detection and quantitation of up to 10 process samples in each test.

View a comparison of all 1D and 2D HCP profiling analyses

2. HCP Antibody Coverage:

Most comprehensive platform to evaluate and compare process specific vs. commercial HCP antibody. Using high resolution, large format 2D fluorescent Western Blot, our HCP Antibody Coverage platform offers high sensitivity detection and accurate quantitation.

3. HCP Impurity Characterization:

Identify and quantify trace amount of HCPs in final product or any step during the process development. By incorporating fractionation at protein level and/or deep scan with AcquireX work flow, we can reliably detect, identify and quantify HCPs at 1-10 ppm level.

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.