cAMP Hunter® Semaglutide
Bioassay
Qualified with Ozempic®
95-0062Y2-00175 (2-Plate Kit)
95-0062Y2-00176 (10-Plate Kit)
Victoza® is the registered trademark of Novo Nordisk A/S
Semaglutide (Ozempic®)
For Treatment of Type II Diabetes mellitus
Mechanism of Action Assay Principle
Source: Sebastião et. al., Front. Endocrinol. (2014); 5, 110
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential
cAMP Hunter® Semaglutide Bioassay Kit
Kit Components
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential 3
Sample data
Semaglutide Bioassay Qualification
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential
Bioassay Workflow
Simple, Homogenous and Rapid Protocol
Plate Read-to-Use Cells Treat with Agonist/ Molecule Read Luminescence
Add Detection Add cAMP
Solution Solution
24 h 30 min 1h 3h
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential
Optimized Semaglutide Dose Curve and Plate Layout
For Assay Qualification
Dilution Scheme Representative Dose Curve Plate Layout
Concentration, Diln
(100%)
ng/mL Factor
3 30000
1 3
0.33 3
0.11 3 20000
RLU
0.056 2
0.037 1.5 10000
0.025 1.5
0.012 2
0
0.0062 2
10 -13 10 -12 10 -11 10 -10 10 -9 10 -8
0.0031 3
[Semaglutide], g/mL
0.00078 4
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential 6
Day-to-Day Repeatability with Semaglutide (Single Analyst)
Day 1 Day 2 Day 3
CH CHO-K1 GLP1R Bioassay CH CHO-K1 GLP1R Bioassay CH CHO-K1 GLP1R Bioassay
Part #: 95-0062Y2 Lot #: 22A2806 Part #: 95-0062Y2 Lot #: 22A2806
Part #: 95-0062Y2 Lot #: 22D1208
30000 30000 30000
20000 20000 20000
RLU
RLU
RLU
10000 10000 10000
0 0 0
10 -13 10 -12 10 -11 10 -10 10 -9 10 -8 10 -13 10 -12 10 -11 10 -10 10 -9 10 -8 10 -13 10 -12 10 -11 10 -10 10 -9 10 -8
[Semaglutide], g/mL [Semaglutide], g/mL [Semaglutide], g/mL
Day 1 Day 2 Day 3 % RSD
S/B 15.8 13.9 16.3 8.3%
EC50,
34.8 29.7 34.1 8.4%
pg/mL
Note: consistent data generated with two different bioassay lots
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential 7
Representative Relative Potency Data (4 Nominal Concentrations)
(Analyst 1 vs Analyst 2)
50% 75% 125% 150%
50% 78.9% 116.7% 149.1%
Analyst 1
47.3% 74.4% 114.8% 151.9%
Analyst 2
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential 8
Semaglutide Bioassay Qualification Data
Nominal Observed RP, Average Average %
RP, % Analyst % RP, % % RSD Recovery
Semaglutide Bioassay Dilutional Linearity
Measured Relative Potency %
1 149.1 200 (50%-150%)
150 1 138.8 143.9 5.4 95.9% R2= 0.9990
150
1 135.8
2 151.9 100
1 130.7
1 116.7 50
`125 119.4 6.4 95.5%
1 115.2 0
2 114.8 0 50 100 150 200
Expected Relative Potency, %
1 108
1 101.1 Parameter Value Specification
100 1 88.1 98.8 7.8 98.8%
1 94.4 Accuracy
(Average % Recovery) 97% 100% +/- 20%
2 102.3
1 73.5 Repeatability 8.7% ≤20%
1 79.0
75 72.8 8.5 97.1% Intermediate Precision ≤8.5% ≤20%
1 64.3
2 74.4 Linearity (R2) 0.9990 ≥0.95
1 47.3
Excellent accuracy, repeatability, intermediate
1 51.1
50 48.9 3.9 97.9% precision, and dilutional linearity
1 50
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently
Confidential Confidential 9
2 47.3
Benefits for “Ready-to-Use” Bioassay Kits
Functional response based on drug MOA
Verified and Qualified with innovator’s marketed drug
Simple protocol; Rapid results
Specific and Sensitive assay
Highly reproducible
Readily Implement with Optimized kit
• Frozen ready-to-assay cells
• Bioassay Detection Reagents
• Cell Plating Reagent
• Dilution Buffer
• Control Agonist
• Tissue Culture-Treated Plates
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential
For More Info, Questions or Technical Support
Web:
Cell-Based Bioassays for Biologics
Technical Support
DRX_SupportUS@[Link]
Drug Discovery & Development Phase-Appropriate Solutions | Our Expertise in Your Hands – Discover Confidently Confidential Rev: 20180410
