JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE) ›› 2024, Vol. 59 ›› Issue (11): 51-63.doi: 10.6040/j.issn.1671-9352.0.2023.504

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Effect of oxidative stress on a cysteine-conjugated antibody-drug conjugate

QU Yaocheng1,2, YAO Xuejing2, SUN Yundong1   

  1. 1. School of Basic Medical Sciences, Shandong University, Jinan 250012, Shandong, China;
    2. RemeGen Co., Ltd., Yantai 264006, Shandong, China
  • Published:2024-11-29

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Abstract: We investigated the effects of oxidative stress on the structure and function of a cysteine(Cys)-conjugated antibody-drug conjugate(ADC). The research object was a Cys-conjugated, c-Met ADC(referred to as c-Met ADC). Firstly, the samples were incubated in different proportions of H2O2 solution for forced oxidation. Then, multiple analytical techniques were used to deeply characterize the structural and functional changes of the oxidized samples. H2O2 induced oxidative stress resulted in Met oxidation on the c-Met ADC antibody backbone and thioether bond cleavage in the linker. Met oxidation further led to a decrease in the thermal stability and neonatal Fc receptor(FcRn)affinity of the c-Met ADC, which were mainly related to Met257 oxidation on the antibody Fc. The cleavage of thioether bonds led to a decrease in the biological activity of the c-Met ADC. Oxidative stress was a key influencing factor on the stability of c-Met ADC, and the Met257 oxidation and thioether bond cleavage caused by it should be strictly monitored as its critical quality attributes in daily release and stability studies. At the same time, it should also be avoided to be exposed to oxidative environments or to mitigate the impact of oxidative stress on its structure and function by optimizing environmental factors.

Key words: antibody-drug conjugate, oxidative stress, forced degradation, methionine oxidation, off-target toxicity

CLC Number: 

  • R917
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