Biopharmaceuticals require products to be free of process-related impurities including Host Cell Protein (HCP) contaminants. These HCPs could be left behind during the purification process from the expression hosts, such as E. coli, insect, or mammalian cells and responsible for serious adverse effects in humans.To examine the presence of residual contamination during the bioprocessing and the final biopharmaceutical product, development of custom polyclonal antibody reagents with maximum coverage and sensitivity against native HCP extracts is required. Please see US FDA Guidelines for Bioanalytical Validation. Rockland’s product portfolio provides well characterized and sensitive generic reagents for immunodetection of host cell proteins as well as multiple customizable options for process-specific HCP antibody development as a service. These options allow flexibility and control to the sponsor for developing early stage through late stage HCP detection reagents.
The Standard method is a process is where the two sets of animals are immunized using high and low molecular weight Host Cell Protein (HCP) extract. After immunization and primary boosting, antisera is collected. The developed anti-HCP antibodies are evaluated to determine coverage of potential HMW and LMW host cell contaminants . The final HCP antibody product is a pool of anti-HMW and anti-LMW antibodies having broad coverage of the HCP antigens in the final antibody reagent. Coverage is demonstrated by 1D western blot analysis or 2D western blot analysis against the HCP extract. HCP-detection assay development is available. To obtain the optimal level of coverage multiple host animals may be utilized. The following species are available; Rabbit, Chicken, Sheep, Goat and Donkey.
Cascade immunization is a method where the animals are boosted using complete Host Cell Protein (HCP) extract. After immunization and primary boosting, antisera is collected. The developed anti-HCP antibodies are purified and used to immunoprecipitate the immunodominant antigens from the HCP extract. The remaining HCP antigens are used to boost the animals in what is called a cascade immunization. This method of immunization has been shown to improve the coverage of the final anti-HCP antibody reagent by driving an immune response and development of antibodies against the weaker immunogens or lower concentration proteins in the HCP. Coverage is demonstrated by western blot or 2D Analysis against the HCP extract. HCP-detection assay development is available. To obtain the optimal level of coverage multiple host animals may be utilized. The following species are available; Rabbit, Chicken, Sheep, Goat and Donkey
One Dimensional SDS-PAGE/Western Blot analysis is performed on test and production bleed s to evaluate the host antibody-mediated immune response to HCP lysate. Both a standard stained gel (Coommassie, Silver or Oriele Stain) and western blot are performed as part of the evaluation. Rockland can customize the testing and analysis methodology to meet your HCP Reagent needs.
Download the HCP Analysis of a Small-Drug Protein in Process Sample Poster
Download the Small HCPs in a 12kDa Protein Drug Analyzed by GeLC-MS/MS Poster
2D-SDS PAGE Analysis Methods For Determining HCP Coverage Using Process Derived anti-HCP Polyclonal Antibodies
Production and Characterization of High Coverage, Process Specific Pichia P. HCP Antibodies
Download the Multi-assay Functional Antibodies in HCP Detection poster
Download the Enhanced Standardized HCP poster
Producing an HCP Antibody: Workflow
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