Method development and validation for determination of antihemolytic activity of eculizumab (Soliris)

Bioanalytical methods are applied at the development and test of drugs as well as at the step of pharmaceutic products issue. Drugs and excipients quality estimation is made by means of precise and reproducible methods. Accuracy and reproducibility of a method is established during validation which is obligatory for medicine registration. The principal task of method validation is experimental evidence of its suitability for the objectives to be achieved. Validation of bioanalytical methods is one of the elements of the whole medicine production validation [1, 2].The aim of research: to validate a method for determination of specific anti-hemolytic activity of eculizumab developed in LCC “IBC Generium”.Materials and methods: eculizumab, antibody-sensitized chicken erythrocytes, complement-containing human serum.Results. We demonstrated the specificity of the method and its correspondence to criteria of accuracy (103.0±1.4)%, robustness (CV – 11.5%), repeatability (CV – (4.9±0.9)%), reproducibility (CV – (3.5±0.4)%), and linearity (k -1.0275; R2 – 0.9975) during validation. The system validity (equipment, materials, analytical operations and analyzed samples) was confirmed for true results obtaining during validation.Results discussion. Experimental evidence of suitability of the method for eculizumab specific activity assessment was obtained in course of validation. The simplicity of the method allows obtaining accurate results in other laboratories. The developed method can be used not only for specific activity of Soliris determination but also for other pharmaceutical substances and drugs based on antibodies specific to human complement C5.

eculizumab, anti-hemolytic activity, validation, specificity, accuracy, linearity, precision, reproducibility