1. Host cell optimization
– Genetically engineered CHO-ks1 cell
– Pre-adaptation to serum-free suspension (minimize lead time by up to 3 months)
– Chemically-defined medium usage
2. Gene sequence optimization
– Codon optimization
– mRNA structure optimization
– Cryptic splice site removal
3. Expression Vector optimization
– Express two genes (heavy chain and light chain) in one vector
– Small sized vector with increased efficiency
– Optimized expression of antibiotics selection marker genes
4. Transfection optimization
– Recombination-based transfection method (proprietary method)
– Up to 12 fold increase in high level expressing stable cell line population
| Population | Method1 | Method2 | Fold increase | |
|---|---|---|---|---|
| Positive cells mean-1 | R3 Peak | 653 | 1,845 | 2.8 |
| Positive cells mean-2 | R6 Peak | none | 6,518 | 10 |
| Positive cells % | R11 population | 19% | 35% | 1.9 |
| Highest Expression cell % | R4 population | 0.7% | 8.4% | 12 |
5. Single cell growth optimization
6. Single cell selection method optimization
– MoFlo-XDP high speed cell sorter usage
– Flow cytometer technologies used to improve sorting of high efficiency cell lines
· Select cells with optimal expression levels with improved ranking consistency
(increase probability of finding optimal producers and select cells)
· Cut cell line/antibody development times – avoid limiting dilution
· Reject poor performers at an early stage based on expression levels in situ
· Pick colonies with accuracy and confidence. Reduce risk of colony disturbance with robotics redesign.
· Improve workflow efficiency with fewer plate handling steps
7. Guaranteed clonality
– Clone Select Imager usage
– Confirm that sorted cell lines are monoclonal (originate from a single cell)
– Eliminate the need for repeated cloning (minimize lead time by 3 months)
– Monitor cell lines from a single cell to a full colony in a 96-well plate
8. Increase productivity via process development
Small-scale production with 1 liter to 10 liter Biostat using Ambr mini-bioreactor
9. Non-GMP production
50 liter Biostat single-use bioreactor for non-GMP production
10.GMP drug production
Osong KBIO Clinical Drug Manufacturing Center’s 200 liter to 500 liter GMP-certified bioreactor usage for large-scale production
11. Purification process development
AKTA Pilot usage
12. Physicochemical analysis
Osong KBIO New Drug Development Center’s analytical instrument (Biacore and Octet) usage
