Optimization of bioprocess conditions improves production of a CHO cell-derived, bioengineered heparin.
Biotechnol J
; 10(7): 1067-81, 2015 Jul.
Article
in En
| MEDLINE
| ID: mdl-26037948
ABSTRACT
Heparin is the most widely used anticoagulant drug in the world today. Heparin is currently produced from animal tissues, primarily porcine intestines. A recent contamination crisis motivated development of a non-animal-derived source of this critical drug. We hypothesized that Chinese hamster ovary (CHO) cells could be metabolically engineered to produce a bioengineered heparin, equivalent to current pharmaceutical heparin. We previously engineered CHO-S cells to overexpress two exogenous enzymes from the heparin/heparan sulfate biosynthetic pathway, increasing the anticoagulant activity â¼100-fold and the heparin/heparan sulfate yield â¼10-fold. Here, we explored the effects of bioprocess parameters on the yield and anticoagulant activity of the bioengineered GAGs. Fed-batch shaker-flask studies using a proprietary, chemically-defined feed, resulted in â¼two-fold increase in integrated viable cell density and a 70% increase in specific productivity, resulting in nearly three-fold increase in product titer. Transferring the process to a stirred-tank bioreactor increased the productivity further, yielding a final product concentration of â¼90 µg/mL. Unfortunately, the product composition still differs from pharmaceutical heparin, suggesting that additional metabolic engineering will be required. However, these studies clearly demonstrate bioprocess optimization, in parallel with metabolic engineering refinements, will play a substantial role in developing a bioengineered heparin to replace the current animal-derived drug.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Heparin
/
CHO Cells
/
Metabolic Engineering
/
Anticoagulants
Limits:
Animals
Language:
En
Journal:
Biotechnol J
Year:
2015
Document type:
Article