Stress Temperature Studies in Small Scale Hastelloy® Drug Substance Containers Lead to Increased Extent of and Increased Variability in Antibody-Drug Conjugate and Monoclonal Antibody Aggregation: Evidence for Novel Oxidation-Induced Crosslinking in Monoclonal Antibodies

Oxidation of polysorbates – An underestimated degradation pathway?

“…These stress conditions include exposure of polysorbates to evaluate the impact of e.g. temperature, iron or transition metals ( Brovč et al, 2020a ; Liu et al, 2022a ; Hvattum et al, 2012 ; Kranz et al, 2019 ; Gopalrathnam et al, 2018 ; Schröter et al, 2021 ; Bensaid et al, 2022 ; Doyle et al, 2019 ; Kranz et al, 2020 ; Klair et al, 2021 ; Prajapati et al, 2022 ), H 2 O 2 ( Kishore et al, 2011a ; Kranz et al, 2019 ; Peters et al, 2022 ), light exposure ( Ha et al, 2002 ; Doyle et al, 2019 ; Doshi et al, 2021a ; Singh et al, 2012 ; Agarkhed et al, 2013 ; Prajapati et al, 2020 , Prajapati et al, 2022 , Larson et al, 2020 ; Lei et al, 2021 ), or 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH) ( Borisov et al, 2015 ; Larson et al, 2020 ; Zhang et al, 2017 ; Schröter et al, 2021 ; Zhang et al, 2018 ), all promoting the formation of radicals resulting in oxidative degradation. As oxidation of polysorbates can occur at: (i) the POE moieties, which can also result in free fatty acids and (ii) the existing double bonds ( Donbrow et al, 1978a ; Zhang et al, 2017 ; Yao et al, 2009 ), it is discussed whether PS80 or PS20 or different subspecies are most susceptible to oxidative degradation (compare Fig.…”

“…(2011), and Klair et al . (2021) to name a few ( Ha et al, 2002 ; Klair et al, 2021 ; Lam et al, 2011 ). Is PS80 more prone to oxidation than PS20?…”

“…Usually, temperature is combined with an additional stressor, like iron ( Liu et al, 2022a ; Doyle et al, 2019 ; Kranz et al, 2019 ; Gopalrathnam et al, 2018 ; Schröter et al, 2021 ; Bensaid et al, 2022 ; Kranz et al, 2020 ; Klair et al, 2021 ; Prajapati et al, 2022 ) or exposure to stainless steel ( Doyle et al, 2019 ; Gopalrathnam et al, 2018 ) to accelerate the oxidation process, since biologicals are in contact with stainless steel upon pharmaceutical manufacturing. Therefore, several groups have stressed PS20 and PS80 in the presence of iron or stainless steel, with different concentrations being applied, ranging from 0.005 mg·L -1 via 0.01 mg·L -1 to 10 mg·L -1 ( Doyle et al, 2019 ; Kranz et al, 2019 ; Bensaid et al, 2022 ; Kranz et al, 2020 ).…”

“…(2021), both determining the iron content by inductively coupled plasma-mass spectrometry (ICP-MS). ( Bensaid et al, 2022 ; Klair et al, 2021 ) Primarily, Fe 2+ SO 4 2- ( Doyle et al, 2019 ; Gopalrathnam et al, 2018 ; Liu et al, 2022a ; Kranz et al, 2019 ; Schröter et al, 2021 ; Kranz et al, 2020 ) or Fe 2+ Cl - 2 ( Bensaid et al, 2022 ; Klair et al, 2021 ) were added, except for Klair et al (2021) and Prajapati et al . (2022), which forced oxidation by Fe 3+ Cl - 3 addition sometimes in combination with other stressors ( Klair et al, 2021 ; Prajapati et al, 2022 ).…”

“…( Bensaid et al, 2022 ; Klair et al, 2021 ) Primarily, Fe 2+ SO 4 2- ( Doyle et al, 2019 ; Gopalrathnam et al, 2018 ; Liu et al, 2022a ; Kranz et al, 2019 ; Schröter et al, 2021 ; Kranz et al, 2020 ) or Fe 2+ Cl - 2 ( Bensaid et al, 2022 ; Klair et al, 2021 ) were added, except for Klair et al (2021) and Prajapati et al . (2022), which forced oxidation by Fe 3+ Cl - 3 addition sometimes in combination with other stressors ( Klair et al, 2021 ; Prajapati et al, 2022 ). The different amounts of iron range from <2 ppb to 15 ppm, depending on the study ( Liu et al, 2022a ; Doyle et al, 2019 ; Kranz et al, 2019 ; Gopalrathnam et al, 2018 ; Schröter et al, 2021 ; Bensaid et al, 2022 ; Kranz et al, 2020 ; Klair et al, 2021 ; Prajapati et al, 2022 ).…”