The document describes TAPBOOST, a breakthrough platform that improves protein production. It works by controlling the protein folding and cellular quality control systems. It enhances yields of target proteins like antibodies, enzymes, and fusion proteins. For example, it increased IL13Rα2-Fc production 2000% in 293T cells and 400% in CHO suspension cells. It also increased yields of antibodies like Humira, Avastin, and DP12 by 600%, 420%, and 730% respectively. The platform works across expression systems like mammalian, bacterial, and microbial to increase both expression levels and solubility of difficult to produce proteins.

1. TAPBOOSTTAPBOOST®®
Breakthrough Platform to Improve Protein Production

2. Misfolded Protein
Protein/Ab Stability
Protein/Ab Secretion
Functional Protein
Immunogenicity
human cell
Polypeptide
Protein Folding
· Suppression of Translation
· Induction/Expression of Chaperones
· Cell suicide
Platform enhances Protein Production of the Target Proteins
by controlling Protein Folding/Cell Quality Control System
RNA
gene
Folded Protein
· Degradation of proteins Degradation
RNA
Protein Folding
Cell Quality Control System
human cell
Total yield of
Biologics/Ab
TAPBOOSTTAPBOOST®®
Protein Folding System and Quality ControlProtein Folding System and Quality Control
SystemSystem

3. Screening System
Step 1: in silico Screening for Protein Domain/Subdomain
Misfolding-Prone
Protein
Degradation
Poor Secretion
Protein Stability
Protein Secretion
Step 3
Protein Stability
and Secretion Test
Develop the technology which controls protein folding/cellular quality control system
specifically for an intended target protein
GOAL
Through the screening,
we discovered the amino
acid sequence with
protein stability and
secretion activity
First Year
Step 2: Library Construction
((TaTargetedrgeted PProteinrotein BoostBoosting Platform)ing Platform)
DiscoveryDiscovery
TAPBOOSTTAPBOOST®®

4. Development of Platform
Step 4: Construction of
Targeting domain
(bind to a target protein)
Discovered sequence
(Effecter Sequence)
Targeting Domain
• Fc binding sequence: 14 combinations
• Antibody binding sequence: 5 combinations
• Target specific sequence: 9 combinations
• Unfolding protein binding sequence: 5 combinations
Flexible Linker / Rigid Liner
• 5 combinations
Effecter Sequence
Signal Sequence
• 4 combinations
Targeted Protein
Second
Year
Step 5: Enhancement of the Production of
Target Protein by
PlatforPlatfor
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
Golgi
ER
TAPBOOSTTAPBOOST®® ((TaTargetedrgeted PProteinrotein BoostBoosting Platform)ing Platform)
Discovery-2Discovery-2

5. Discovered sequence
Sequence Optimization
Deletion of
the sequence
(49 deletion mutants)
Minimum sequence
(9 amino acid sequence)
Step 6: Identification of the Minimum Sequence
Various mutants of the sequence
(49 mutations)
Step 7: Formulation of the Sequence
X1 X2 X3 X4 X5 X6 X7 X8 X9
Formulation of the Amino Acid Sequence
((TaTargetedrgeted PProteinrotein BoostBoosting Platform)ing Platform)
Discovery-3Discovery-3
TAPBOOSTTAPBOOST®®

6. Model Protein; IL-13 receptor α2 (IL13Rα2)-Fc fusion protein
CHALLENGE: The expression levels of
IL13Rα2 are poor, and the protein is
prone to misfolding and aggregation.
Cell
IL13Rα2-Fc
ERNu
• Disease Relevance: a key mediator of allergic inflammation
Aggregation of
IL13Rα2-Fc
Only properly folded
IL13Rα2-Fc secrets
• Fc fusion protein with sIL13Rα2 (sIL13Rα2-Fc) has been developed for the
treatment of asthma
• Production of sIL13Rα2-Fc using human cells is inefficient due to its
instability
• Soluble IL13Rα2 (sIL13Rα2) blocks the signal from IL-13 as a decoy
receptor
Application: IL13RApplication: IL13Rαα2-Fc Protein Case2-Fc Protein Case
StudyStudy
TAPBOOSTTAPBOOST®®

7. 0
500
1000
1500
2000
2500
0
20
40
60
80
100
120
Secreted
IL13Rα2-Fc
IL13Rα2-Fc
Inside Cells
Secreted IL13Rα2-Fc IL13Rα2-Fc Inside Cells
IL13Rα2-Fc
FoldIncrease(%)
FoldIncrease(%)
IL13Rα2-Fc
Case Study - IL13RCase Study - IL13Rαα2-Fc2-Fc
ProteinProtein
TAPBOOSTTAPBOOST®®
enhances the secretion
of IL13Rα2-Fc Protein
TAPBOOSTTAPBOOST®®
0
500
1000
1500
2000
2500
0
20
40
60
80
100
120
>20x
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®

8. ER
Targeted Protein
Targeting Domain was designed
to bind only to Protein B
Secreted
Target Protein
(A or B)
A A A B B B B BTarget Protein
Platform: Target Protein SpecificityPlatform: Target Protein SpecificityTAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
specifically enhances the secretion of the
target proteins
TAPBOOSTTAPBOOST®®
B

9. enhances
the production of all three
tested antibodies
Antibody Production
Detection: Western Blot Assay using anti-hIgG antibody
Platform: Antibody ExamplesPlatform: Antibody ExamplesTAPBOOSTTAPBOOST®®
Western
(anti-hIgG Ab)
TAPBOOSTTAPBOOST®®
Target Antibody
anti-IL8 Ab
(DP12)
anti-TNFα Ab
(Humira)
anti-VEGF Ab
(Avastin)
antibody Brand Name
Production
Increase
(produced protein)
anti-IL8 Ab DP12 6.4 +/- 0.89
anti-TNFα Ab Humira 6.3 +/- 0.33
anti-VEGF Ab Avastin 3.5 +/- 0.27
TAPBOOSTTAPBOOST®®

10. Antibody binding affinity for its corresponding epitope
Detection: ELISA using rVEGF, rTNFα, or rIL-8 for Avastin, Humira, or DP12, respectively
Platform: Antibody ExamplesPlatform: Antibody ExamplesTAPBOOSTTAPBOOST®®
Antibodies produced
by
hold activity
TAPBOOSTTAPBOOST®®
anti-TNFα Ab
(Humira)
anti-VEGF Ab
(Avastin)
anti-IL8 Ab
(DP12)
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
0
1
2
3
4
5
6
7
8
0
1
2
3
4
5
6
7
8
FoldIncrease
Avastin
FoldIncrease
FoldIncrease
Humira DP12
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
antibody Brand Name
Production
Increase
(produced protein)
Production Increase
by ELISA
anti-VEGF Ab Avastin 3.5 +/- 0.27 4.2 +/- 0.19
anti-TNFα Ab Humira 6.3 +/- 0.33 6.0 +/- 0.82
anti-IL8 Ab DP12 6.4 +/- 0.89 7.3 +/- 0.56

11. substantially
enhances the production of
Factor VIII-Fc
Platform: Enzyme ExamplePlatform: Enzyme ExampleTAPBOOSTTAPBOOST®®
Foldincrease
FVIII Production
0
2
4
6
8
10
12
14
16
18
20
FVIII-Fc
Foldincrease
FVIII Activity
0
2
4
6
8
10
12
14
16
18
20
22
24
FVIII-Fc
Target Protein
Production Increase
(produced protein)
Production Increase
by Activity
FVIII-Fc 19.2 +/- 0.47 21.3 +/- 0.23
TAPBOOSTTAPBOOST®®
0
2
4
6
8
10
12
14
16
18
20
TAPBOOSTTAPBOOST®®
0
2
4
6
8
10
12
14
16
18
20
22
24
TAPBOOSTTAPBOOST®®

12. 0
2
4
6
8
10
12
Foldincrease
TNFRI-Fc (IgG1)
TNFα Binding Ability (ELISA)
TNFRI-Fc (IgG4)
Platform: Fc Fusion of hIgG4Platform: Fc Fusion of hIgG4TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
enhances the production
of both types of Fc fusion protein
TAPBOOSTTAPBOOST®®

13. IL13Rα2-FcTNFRI-Fcα1AT-Fc α1ATMT-Fc
Original Secretion of
Targeted Protein
Effect of
Maximum
Effect
may “rescue” problem proteins from
misfolding and maximize secretion levels
works on difficult proteinsworks on difficult proteinsTAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®

14. Protein Aggregation Experiment
conducted by Major Pharma Partner in Pilot Project
• PD group wanted to know whether can also results in lower aggregation.
CharacterizationCharacterization
TAPBOOSTTAPBOOST®®
reduces the aggregation
of produced proteins
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®

15. Target Protein
(Conditioned Media)
FoldIncrease(%)
IL13Rα2-Fc
IL13Rα2-Fc
Protein Production in CHO Suspension conducted by CRO
0
50
100
150
200
250
300
350
400
450
enhances the production in CHO
Suspension Expression System
Transition from 293T to CHO suspensionTransition from 293T to CHO suspension
systemsystem
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®

16. PKCActivity(OD450-BG)
AT1 receptor
-AT1 receptor
Angiotensin II (100nM) 5min 10min 15min 30min
Angiotensin II receptor type 1
(AT1 receptor)
AT1 receptor
-AT1 receptor
+ Angiontensin II
Application for Screening R&D – GPCR proteins (POC)Application for Screening R&D – GPCR proteins (POC)
enhances the expression of difficult-to-express
proteins such as GPCR proteins
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®

17. Soluble protein (SP)
Inclusion Body (IB)
bacteria
Bacterial Genome
plasmid
We customized platform for bacterial
expression system by using a proprietary algorithms
and intensive search for specific amino acid
sequences involved in protein folding.
Platform
Test Protein A
Test Protein B
Test Protein C
SP IBTotal SP IBTotal
SP IBTotal SP IBTotal
SP IBTotal SP IBTotal
Application for Microbial ExpressionApplication for Microbial Expression
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
increases both expression and solubility of the
target protein in bacteria
TAPBOOSTTAPBOOST®®

18. Advantages of
SummarySummary
Increased Productivity byTarget proteins
TNFRI-Fc (Enbrel) 480%
Factor IX-Fc 320%
Factor FVII-Fc 1000%
IL13Rα2-Fc protein 2000%
Factor VIII-Fc 2100%
Humira
Avastin
DP12
600%
420%
730%
293T monolayer System
293T Suspension System
IL13Rα2-Fc protein 1100%
CHO Suspension System
IL13Rα2-Fc protein 400%
* Optimization process is in progress
*
*
(%)TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®
• Any kind of protein can be expressed by platform
• Easy implementation into your current bioprocess
• Combine with any other technology for synergistic effect
TAPBOOSTTAPBOOST®®
TAPBOOSTTAPBOOST®®