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Stability of monoclonal antibodies after simulated subcutaneous administration

Published:March 11, 2021DOI:https://doi.org/10.1016/j.xphs.2021.03.007

      Abstract

      Changes in the environment from the drug product to the human physiology might lead to physical and/or chemical modifications of the protein drug, such as in vivo aggregation and fragmentation. Although subcutaneous (SC) injection is a common route of administration for therapeutic proteins, knowledge on in vivo stability in the SC tissue is limited. In this study, we developed a physiologic in vitro model simulating the SC environment in patients. We assessed the stability of two monoclonal antibodies (mAbs) in four different protein-free fluids under physiologic conditions. We monitored protein stability over two weeks using a range of analytical methods, in analogy to testing purposes of a drug product. Both mAbs showed an increase of protein aggregates, fragments, and acidic species. mAb1 was consistently more stable in this in vitro model than mAb2, highlighting the importance of comparing the stability of different mAbs under physiologic conditions. Throughout the study, both mAbs were substantially less stable in bicarbonate buffers as compared to phosphate-buffered saline. In summary, our developed model was able to differentiate stability between molecules. Bicarbonate buffers were more suitable compared to phosphate-buffered saline in regards to simulating the in vivo conditions and evaluating protein liabilities.

      Graphical abstract

      Keywords

      Abbreviation:

      AF (artificial fluid), AF+HA (artificial fluid + hyaluronan), BY (brown-yellow), CE-SDS (capillary electrophoresis - sodium dodecyl sulfate), cIEF (capillary isoelectric focusing), CO2 (carbon dioxide), FAL (Float-A-Lyzer), HA (hyaluronan), HMWS (high-molecular weight species), HP-SEC (high performance - size-exclusion chromatography), LMWS (low-molecular weight species), LO (light obscuration), mAb (monoclonal antibody), NTU (nephelometric turbidity units), PBS (phosphate-buffered saline), pI (isoelectric point), RM (reference material), SAL (Slide-A-Lyzer), SbVP (subvisible particle), SC (subcutaneous)
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