Pascale Bardo-Brouard1, Victoire Vieillard2, Tala Shekarian3, Aurélien Marabelle4, Alain Astier2, Muriel Paul2. 1. Department of Pharmacy, Henri Mondor University Hospitals, AP-HP, 51 Avenue du Maréchal de Lattre de Tassigny, 94010 Créteil, France. Electronic address: pascale.bardo@aphp.fr. 2. Department of Pharmacy, Henri Mondor University Hospitals, AP-HP, 51 Avenue du Maréchal de Lattre de Tassigny, 94010 Créteil, France. 3. CNRS UMR5286, INSERM U1052, Cancer Research Center of Lyon, Leon Berard Cancer Center, Lyon, France. 4. CNRS UMR5286, INSERM U1052, Cancer Research Center of Lyon, Leon Berard Cancer Center, Lyon, France; Cancer Immunotherapy Program Drug Development Department (DITEP), INSERM U1015, Gustave Roussy, 114, rue Édouard-Vaillant, 94805 Villejuif Cedex, France.
Abstract
PURPOSE: Well-documented stability data of monoclonal antibodies are generally missing. That is why we studied the physicochemical and biological stability of undiluted ipilimumab (IPI) in glass vial (5 mg/ml) over 28 d after opening, stored at 4 °C and 25 °C. METHOD: A stressed study (60 °C) was performed to validate our analytical methods as 'stability indicating'. The different methods used were turbidimetry, dynamic light scattering (DLS), second-derivative ultraviolet and chromatographic methods as size-exclusion chromatography (SEC) and cation-exchange (CEX). Biological characterisation was performed by an in vitro functional binding inhibition bioassay. RESULTS: We demonstrated that ipilimumab in opened vials stored at 4 °C and 25 °C remained stable for at least 28 d. No physical, chemical or structural instability was found. No aggregation was observed by turbidimetry, SEC and DLS. Hydrodynamic diameters remained unchanged, as chromatographic profiles in CEX and thermal aggregation curves. Functionally, the ability of IPI to antagonise CTLA-4/B7.2 binding remained stable over 1 month at 4 °C. CONCLUSION: These results indicate that unused residues of IPI in their original vials can be safely kept up to 28 d, following good manufacturing procedures, allowing re-use for another patient or in case of cold-chain rupture.
PURPOSE: Well-documented stability data of monoclonal antibodies are generally missing. That is why we studied the physicochemical and biological stability of undiluted ipilimumab (IPI) in glass vial (5 mg/ml) over 28 d after opening, stored at 4 °C and 25 °C. METHOD: A stressed study (60 °C) was performed to validate our analytical methods as 'stability indicating'. The different methods used were turbidimetry, dynamic light scattering (DLS), second-derivative ultraviolet and chromatographic methods as size-exclusion chromatography (SEC) and cation-exchange (CEX). Biological characterisation was performed by an in vitro functional binding inhibition bioassay. RESULTS: We demonstrated that ipilimumab in opened vials stored at 4 °C and 25 °C remained stable for at least 28 d. No physical, chemical or structural instability was found. No aggregation was observed by turbidimetry, SEC and DLS. Hydrodynamic diameters remained unchanged, as chromatographic profiles in CEX and thermal aggregation curves. Functionally, the ability of IPI to antagonise CTLA-4/B7.2 binding remained stable over 1 month at 4 °C. CONCLUSION: These results indicate that unused residues of IPI in their original vials can be safely kept up to 28 d, following good manufacturing procedures, allowing re-use for another patient or in case of cold-chain rupture.