Cell Line Development and Cell Analysis Technologies

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