Evaluation of the pattern and kinetics of degradation of adalimumab using a stability‐indicating orthogonal testing protocol

Abstract

Forced degradation studies are crucial for the evaluation of the stability and biosimilarity. Here, adalimumab was subjected to oxidation, pH, temperature, agitation and repeated freeze-thaw in order to generate all possible degradation products. An orthogonal stability-indicating testing protocol comprising SE-HPLC, RP-HPLC, TapeStation gel electrophoresis, dynamic light scattering (DLS), and functional receptor binding assay was developed and validated. The assay protocol was used for the assessment of the pattern and kinetics of aggregation/degradation of adalimumab. SE-HPLC and DLS were used to show the formation of aggregates/fragments of adalimumab under nondenaturing conditions. TapeStation electrophoresis was performed under denaturing conditions to reveal the nature of aggregates. Results of the receptor binding assay agreed to those of SE-HPLC and DLS which indicated that it can be used as an activity-indicating assay for adalimumab. RP-HPLC demonstrated excellent selectivity for adalimumab in the presence of its oxidized forms. The kineticsof degradation was studied in each case and the results showed that it followed the first-order reaction kinetics. Correlation between the results supported the quality assessment of the tested product in industrial and clinical settings. This orthogonal protocol is a useful tool in stability assessment of monoclonal antibodies and a key criterion for the biosimilarity assessment.

Stability of Monoclonal Antibodies as Solid Formulation for Auto-Injectors: A Pilot Study

“…Normally, “the least stable domain is involved in the aggregation process” [ 13 ], which means that for ADA it would be the C H 2 domain. In any case, it has been demonstrated that enthalpy reductions are related to losses of native antibody structure, which sometimes has been associated with aggregation events [ 21 , 22 ]. The hydrodynamic size and ζ potential of the ADA dilutions were also determined.…”

“…From these results, it is possible to conclude that temperature-stress conditions for ADA translate into an enthalpy decrease with no significant changes in transition midpoints when analyzed by nano-DSC. It has already been reported that temperature-stress conditions induce ADA aggregates of different MW and solubility [ 22 , 24 ]. Wu et al stated that aggregation is temperature dependent, having an Arrhenius regime for T ≥ T m1 (C H 2 transition midpoint) or a non-Arrhenius regime for T < T m1 [ 25 ].…”

Investigation of the Physicochemical and Biological Stability of the Adalimumab Biosimilar CT-P17

“…Biological products tend to be less stable than small molecules and, importantly, subtle changes induced by environmental stress can significantly affect their therapeutic properties [25]. In addition, monoclonal antibodies are frequently formulated at high concentration levels, potentially leading to aggregate formation and resultant impurities [27]. Further, stability testing also allows recommended storage conditions to be established [25].…”