Prevention |
Inhibit enzyme overexpression |
Knocked down the expression
of thioredoxin (Trx) in CHO cells |
Koterba, Borgschulte, & Laird,
2012 |
|
Inhibit enzyme activity during cell culture and cell harvest
|
Induce Ethylenediaminetetraacetic acid (EDTA) to inhibit hexokinase
activity and reduce the formation of glucose-6-P;
Induce Cu2+, Zn2+,
Hg2+ and Co2+ to inhibit Trx
activity;
Induce Aurothioglucose (ATG) and aurothiomalate (ATM) to inhibit
thioredoxin reductase activity
|
Kao, Hewitt, Trexler-Schmidt, & Laird, 2010;
Trexler-Schmidt et al., 2010
|
|
Reduce enzyme amount for disulfide bond reduction during cell culture,
cell harvest and clarified bulk (CB) storage
|
Induce O2, H2O2 and
cysteine as enzyme competitors;
|
Du et al., 2018; Mun et al., 2015; Chung et al., 2017;
Handlogten, Wang, & Ahuja, 2020
|
|
|
Control centrifuge force during harvest to minimize cell lysis;
Control depth filtration pressure to minimize cell lysis;
|
Trexler-Schmidt et al., 2010; O’Mara et al., 2019
|
|
Slowdown enzyme reaction rate during cell harvest and CB storage |
Lower pH and temperature |
Xie et al., 2016; Hutterer et al., 2013; Liu,
Nowak, Shao, Ponniah, & Neill, 2016 |
|
Reduce reaction time |
Shorten CB storage time |
Mun et al., 2015;
O’Mara et al., 2019 |
Rescue |
Reoxidize reduced mAb during Protein A chromatography |
Introduce redox wash to reoxidize reduced mAb |
Tan et al., 2020; Tang
et al., 2020 |