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Pharmacokinetic Assay Development

The assessment of drug efficacy and safety is of major concern in the pre-clinical and clinical testing phases of biologics drug development. As suggested by the “LADME” scheme, the distribution, metabolization and elimination of a pharmacologically active substance needs to be monitored. PK assays are used to determine the concentration of the drug in a relevant sample (e.g. patient serum) at given times. Regulatory authorities (the EMA and FDA) have released several guidelines for how these issues should be addressed using in vitro studies. 

 

Assay Format 

Sandwich ELISA principle. Blue: monoclonal anti-ID (biotinylated or enzyme-conjugated); orange: target drug antibody (analyte); anti-ID conjugation enables assay readout

The sandwich ELISA is BioGenes' preferred approach of PK assays. This means that the therapeutic drug is successively captured and detected by two monoclonal antibodies, in a layered fashion.

Monoclonal anti-IDs are routinely used as capture and detecting reagents in pharmacokinetic (PK) assays for the quantification of antibody drugs in patient serum.

Development Strategy

Our experts understand the difficulties and complexities that need to be addressed in your studies and will help you determine the optimal approach for PK and immunogenicity testing.  

Phase 1 - Expert consultation

  • Definition of the project´s aim
  • Thourough discussion of antigen characteristics and project requirements

 

Phase 2 - Antigen preparation and immunization

  • Antigen preparation (optional Fab or F(ab)2 Fragmentation) and characterization
  • Immunization of mice according to BioGenes’ enhanced immunization protocol or customized protocols (upon request)
  • Titer determination & selection of mice for hybridoma generation
  • Optional: Prolongation of immunization schedule to achieve appropriate titres

Phase 3 - Cell fusion and hybridoma screening

  • Generation and selection of hybridoma cell lines
  • Screening for IgG producing hybridomas by ELISA including competitive ELISA with 
    drug antigen as well as ELISA screening with human, rat or cynomolgus sera

Phase 4 - Cloning

  • At least two cloning cycles and screening for relevant hybridomas
  • Delivery of cell culture supernatants for each clone for in-house testing by the customer
  • Characterization of selected clones (isotype determination and mycoplasma testing)

Phase 5 - Pair search and antibody production

  • Monoclonal antibodies will be used for the antibody pair search (capture and detector)
  • Antibody pair characterization of 5-6 pairs in different sera (human, rat, cynomolgus)
  • Large-scale antibody production of two selected clones
  • Preparation of the detector antibody-biotin conjugate

Phase 6 - ELISA development

  • ELISA set-up based on selected antibody pair 
  • Testing and optimization of the ELISA parameters 
  • Testing of the ELISA accuracy and precision 

Phase 7 - Method transfer

  • Transfer of the antibodies or the assay method to FyoniBio or the customer’s preferred CRO 
  • Opt. consultation with hands-on training 

Approximate project duration: 12-18 months

Pooled Scientific Expertise

To provide you with an all-inclusive service, BioGenes works closely together with FyoniBio. By combining our scientific expertise, we can offer you an all-in-one package ranging from custom antibody and assay development to ELISA validation and the analysis of pre-clinical and clinical samples according to GCLP guidelines. However, customers can also choose to independently obtain the individual services from either company alone.

 

Contact 

If you need more information, a specific offer 
or want to talk to an expert

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