Development and Validation of an Enzyme-Linked Immunosorbent Assay to Measure Free Eculizumab Concentration in Serum

Abstract

Aim: Eculizumab is a monoclonal antibody toward C5 fraction of the complement system. It is approved to treat paroxysmal nocturnal hemoglobinuria and atypical hemolytic uremic syndrome. To perform pharmacokinetic studies and therapeutic drug monitoring, a validated assay is required. Materials & methods: An indirect ELISA with recombinant human C5 sensitized microtiter plates were developed. Results: The assay allows the measurement of free eculizumab concentration in human serum. The LOD, LLOQ and ULOQ were 0.091, 0.25 and 82.35 mg/l, respectively. The assay meets EMA and US FDA guidelines criteria for the validation of a ligand-binding assay. Conclusion: This method is validated and can be used in PK and PK-PD studies as well as to perform therapeutic drug monitoring of free eculizumab.

Keywords::

• biodrug
• biologic
• biopharmaceutical
• eculizumab
• ELISA
• monoclonal antibody
• pharmacokinetics
• pharmacokinetic–pharmacodynamic
• therapeutic drug monitoring

Supplementary Data

Financial & competing interests disclosure

The study was supported by the French National Research Agency under the programme ‘Investissements d’avenir’ Grant Agreement LabEx MAbImprove: ANR-10-LABX-53. Measurement of serum eculizumab concentrations was performed within the platform Pilot centre for Therapeutic Antibodies Monitoring (PiTAM/CePiBAc). PiTAM/CePiBAc is cofinanced by the FEDER European Union programme (Fonds Européen de développement régional—European funding for regional development).

D Ternant has given lectures for Amgen and Sanofi. G Paintaud reports grants received by his research team from Novartis, Roche Pharma, Genzyme, MSD, Chugai and Pfizer, outside of the submitted work. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

The study was supported by the French National Research Agency under the programme ‘Investissements d’avenir’ Grant Agreement LabEx MAbImprove: ANR-10-LABX-53. Measurement of serum eculizumab concentrations was performed within the platform Pilot centre for Therapeutic Antibodies Monitoring (PiTAM/CePiBAc). PiTAM/CePiBAc is cofinanced by the FEDER European Union programme (Fonds Européen de développement régional—European funding for regional development).