Are you looking for a reliable and customized HCP assay development process for your project? BioGenes has been developing process-specific HCP ELISAs since 1999. With over 400 projects for various cell lines, BioGenes has become a leading global specialist in this field. Our highly qualified scientists and a proactive project management team enable BioGenes to offer a complete portfolio of services. Our experience has resulted in a state-of-the-art specific HCP ELISA development approach that is appreciated by our customers worldwide.

It's important to note that the ELISA development process is time-consuming and typically takes between 1 to 1.5 years to complete. To ensure a smooth and efficient process, BioGenes recommends initiating your project at an early stage, preferably when the proof of concept has been achieved. 

Project Steps


Our typical customized HCP assay development workflow includes the following steps:

Phase 1 - Expert Consultation

  • Definition of the project's aim 
  • Thorough discussion of MOCK material characteristics 
  • Definition of immunization strategy (protocol, species, animal number)
  • Extensive discussion between BioGenes’ scientists, project manager and customer to provide a specific offer including defined work phases

Phase 2 - Antigen Characterization

  • Evaluation whether antigen material represents required HCP composition (e.g. harvest of production process)
  • 2D DIGE comparison of the mock HCP sample with an actual process sample to ensure suitability of the mock sample
  • Optional: preparation of low molecular weight (LMW) HCP fraction to ensure antibody generation against smaller, less immunogenic proteins

Phase 3 - Antibody Generation

  • Immunization of two different species with total mock-HCP and LMW mock-HCP in parallel
  • Affinity purification using the immobilized HCP material (total IgG purification by protein A/G only in case of antibody loss)
  • Comparison of antibodies obtained from two different species in a preliminary ELISA set-up to determine sensitivity of HCP detection
  • Estimation of HCP log-reduction over consecutive DSP steps, determination of sufficient dilution linearity and exclusion of cross-reactivity towards drug substance
  • Selection of most suitable species for 2nd immunization cycle
  • Prolonged immunization period (up to 7 months) for increased affinity of HCP antibodies
  • Preparation of final bulk antibody including affinity purification, biotinylation of detector antibody and aliquot preparation of capture and detector antibodies

Phase 4 - Assay setup, qualification and reagent characterization

  • ELISA set-up and subsequent optimization (accuracy, precision and dilution linearity) including testing of different matrices.
  • System suitability test including comprehensive acceptance criteria (e.g. criteria for HCP standard curve and independent assay control)
  • Coverage determination by different orthogonal methods (2D Western blot, Immunoaffinity Chromatography (IAC) + 2D-DIGE and IAC + LC-MS)

Phase 5 - Pre-Validation

  • Pre-validation of the ELISA according to the ICH-Guideline Q2 (R1): “Validation of Analytical Procedures: Text and Methodology” (CPMP/ICH381/95), November 2005
  • Preparation of a pre-validation protocol before and corresponding report after the study
  • Determination of specificity, accuracy, repeatability, intermediate precision, LOD, LOQ and working range
  • Optional: robustness testing

Phase 6 - Transfer of ELISA method into kit production

  • Preparation and testing of the kit’s component panel, documentation for the production process (SOP) and ELISA instruction leaflet
  • Production of ELISA kits for real-time testing and / or pre-validation
  • Real-time stability testing of ELISA kits

Phase 7 - Kit production/deliverables

  • Kit production – different lot sizes available on request
  • Release testing by QC including CoA
  • Temperature controlled shipment of ready-to-use kits

Latest Publications


  • Preparation of low molecular weight (LMW) fraction and total HCPs for immunization
  • Flexible immunization protocols with different animal species
  • Antibody purification tests of a representative antiserum pool
  • Assay pre-validation is performed by established assay parameters according to ICH guideline Q2 (R1)
  • Reagent production for long-term availability


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