Biological stability of Pembrolizumab after dilution and storage in 0.9% NaCl infusion polyolefin bags
5 October 2022
A. Acramel1,2, G. Le Guyader3,4, K. Avrillon4, L. Guyonnet5, J. Denizeau6,7, L. Escalup1, R. Desmaris1, E. Piaggio6,7, C. Guerin5, C. Sedlik6,71 Institut Curie, PSL Research University, Pharmacie Department, F-75005 Paris, France
2 Université Paris Cité, CiTCoM, UMR8038 CNRS, U1268 Inserm, F-75006 Paris, France
3 APHP, Hôpital Henri Mondor, Pharmacie, F-94090 Créteil, France
4 Centre Hospitalier Intercommunal de Créteil, Pharmacie, F-94000 Créteil, France
5 Institut Curie, Cytometry Platform, F-75005 Paris, France
6 Institut Curie, PSL Research University, Translational Research Department, F-75005 Paris, France
7 Inserm U932, F-75005 Paris, France
Objective
Pembrolizumab is an anti-PD1 human IgG4 monoclonal antibody used for the treatment of various types of cancer. According to the supplier’s recommendations, the diluted solution of pembrolizumab at 1 mg/ml in a 0.9% saline solution can be stored for at least 1 week at 4°C or at room temperature (RT). Recently, we demonstrated by convergent methods that solutions at 1 mg/ml and 4 mg/ml could be stored for 2 weeks at 4°C and 1 week at RT without physicochemical alterations. However, only solutions at 4 mg/ml were stable after 30 days of storage at 4°C. The aim of this work is to complete these data with a biological stability study of Pembrolizumab solutions diluted at 1 and 4 mg/ml.
Methods
Pembrolizumab solutions were diluted in 0.9% NaCl at concentrations of 1 and 4 mg/ml in infusion polyolefin bags. These bags were prepared sequentially and stored at 4°C or 25°C to carry out all the biological tests on the same day for all the samples. Physicochemical analyzes (ion and size exclusion chromatography, dynamic light scattering, UV and fluorescence spectroscopy) were carried out to ensure the compliance of these solutions.
The tests were performed in triplicate from 3 batches of Pembrolizumab. The biological stability of the samples was assessed on one hand using a commercial ELISA test which measures the quantity of Pembrolizumab present in the bags and on the other hand by flow cytometry which detects the binding of the antibody to the cell surface. The in vitro cultured Jurkat-PD1 cell line was used for flow cytometry assay using secondary antibodies specific for human Ig kappa. The samples stored at 4°C were studied on D0, D14 and D28; the samples stored at 25°C on D0 and D7.
Results
Compared with the functional activity at D0, no significant difference was observed between the different samples at the different concentrations and storage conditions studied. The binding capacity of Pembrolizumab on its target is stable at 4°C for 28 days and 7 days at room temperature at 1 mg/ml and 4 mg/ml.
Discussion-Conclusion
The biological stability of Pembrolizumab indicates that the binding of Pembrolizumab to PD1-expressing cells is not altered under the conditions evaluated here. These results suggest that the mechanisms involved in the clinical effect are conserved and therefore add more stability data on Pembrolizumab. The stability of preparations made in anticancer production unit could therefore be reconsidered.