• Objective Optimization of non-reduced CE-SDS analysis of mAb1

• Factors Four continuous variables and two discrete variables

• Responses IgG Purity and resolution (between intact mAb and HHL)

• Design Fractional factorial

• Objective Assess ruggedness of SEC-HPLC method for MAb1

• Factors Four discrete variables

• Responses % High Molecular Weight Impurity
• Design Staggered design

Source Variance % of Total Variance Standard Deviation

Multi-Vari Chart for %HMW by MP_Lot - Analyst_Day
Analyst_Day -2.57475e-008* 0.00% 0.0000000
1 2
1, 1 1, 2
± ± ° 0.01 60
MP_Lot
1
HPLC -1.69324e-007* 0.00% 0.0000000
2
0.01 55 Column 0.0000001 8.51% 0.0002711

0.01 50 MP_Lot -8.30739e-008* 0.00% 0.0000000

0.01 45 Analyst_Day*HPLC 0.0000001 10.81% 0.0003056

%HMW
0.01 40 Analyst_Day*Column -8.16774e-008* 0.00% 0.0000000
2, 1 2, 2
0.01 60
Analyst_Day*MP_Lot -3.91136e-008* 0.00% 0.0000000
0.01 55
HPLC*Column -3.31761e-008* 0.00% 0.0000000
0.01 50
HPLC*MP_Lot 0.0000002 23.51% 0.0004506
0.01 45
• To identify critical • To understand the
Assay Optimization

Assay Robustness

• To evaluate
Assay Ruggedness

Column*MP_Lot -8.38011e-008* 0.00% 0.0000000

parameters of the impact of variation in uncontrolled, normal 0.01 40
1 2
analytical method. test parameter variation due to Column
Error 0.0000005 57.17% 0.0007027
• To establish set points settings upon the typically discrete Panel variables: Analyst_Day, HPLC Total 0.0000009 N/A 0.0009294
and acceptable performance of the factors such as
ranges for method test method. different analysts,
• Objective Assess robustness of potency ELISA method for MAb1
parameters. labs, equipment, and
days. • Factors Three continuous variables
Accuracy and precision at 50%, 100% and 150% of target
• Responses 𝜎𝑀𝑆 = 𝑅𝑒𝑝𝑒𝑎𝑡𝑎𝑏𝑖𝑙𝑖𝑡𝑦 2 + 𝑅𝑒𝑝𝑟𝑜𝑑𝑢𝑐𝑖𝑏𝑖𝑙𝑖𝑡𝑦 2
concentration
• Design Full factorial

𝑃ൗ 𝑅𝑎𝑡𝑖𝑜 = 6 × 𝜎𝑀𝑆 ∗ 100

𝑇 𝑈𝑆𝐿 − 𝐿𝑆𝐿

𝑃ൗ 𝑅𝑎𝑡𝑖𝑜 = 3 × 𝜎𝑀𝑆
𝑇 ∗ 100
𝑈𝑆𝐿 − 𝑇𝑎𝑟𝑔𝑒𝑡

Assay Level of readiness DoE Studies

Assay parameters set point identified and
Residual DNA by PCR • Ruggedness
ranges defined
• Robustness
Relative Binding by ELISA Assay parameters set point identified
• Ruggedness

Peptide Mapping Preliminary protocol defined

Purity by CE-SDS Preliminary protocol defined • Optimization

• Robustness
Charge Variant by icIEF Preliminary protocol defined • Ruggedness

Monomer Content by SEC Preliminary protocol defined

www.postersession.com