7-7-7: Low Endotoxin & Low Aggregate Antibody Production

Production of antibodies that have low endotoxin (<0.5 EU/mg) and aggregates (>95% monomer) levels at 1-100 mg scale, suitable for both in vitro and in vivo studies.

Protein Sciences: HTP Production of High-Quality Vanilla Antibodies

WuXi Biologics’ industry-leading R&D-scale custom protein production group generates high-quality proteins and antibodies to meet your schedule and requirements. No project is too large or small. As part of these services, we offer expedited 777 mammalian transient production for low endotoxin and low aggregate antibodies in just 3 weeks, suitable for both in vitro and in vivo studies.

777 Antibody Production

7 days for Gene Synthesis • AI Codon Optimization 7 days for Protein Expression • Average mAb Titer 900 mg/L 7 days for Purification and QC • >95% High Purity • Low Endotoxin

High-quality antibody is good for most in vitro and in vivo studies Low aggregation (>99% monomer) Low endotoxin (<0.5 EU/mg)

7

7

Many scales are available (yields are dependent on molecules) 20 mL (3-10 mg)

These timelines represent the typical turnaround time for a regular monoclonal antibody. More complex molecules such as bispecific antibodies, Fc-fusion proteins, and other recombinant proteins usually have longer timelines.

7

100 mL (15-100 mg) 200 mL (30-200 mg) 500 mL (100-500 mg)

FREE Cloning & DNA Preparation

Case Study: Regular IgG Antibodies at 20 mL Each

Below is an example of 170 IgG antibodies from early discovery before any optimization or process development. The molecules behave quite differently which is a characteristic of early-stage leads. This case is a true representation of WuXi Biologics Protein Sciences’ capabilities. We achieved average 225 mg/L yield after 2-step automated purification while maintaining endotoxin <0.1 EU/mg without endotoxin removal step.

14

0.000 0.020 0.040 0.060 0.080 0.100 0.120 0.140 0.160

1.2

100

99.8%

12

1

99.5

10

0.8

99

0.7 g/L

8

0.6

98.5

6

4.5 mg

0.4

98

4

97.5

0.2

2

0.015 EU/mg

0

97

0

777 Service Details:

Service Item

Deliverables

Duration

QC

Gene synthesis, cloning and plasmid prep all included

20 mL CHO/HEK293 500 mL CHO/HEK293

3-10 mg, average 4.5 mg after 2-step purification 100-500 mg

3 weeks

A280, Caliper (R/NR) or SDS-PAGE (R/NR), SEC-HPLC, and endotoxin (<0.5 EU/mg) SEC-HPLC 7.922

1.dx

100 150 200 250 300 350 400

3 weeks

Additional QC

RP-HPLC, bioburden, DSF, DSC, SPR, LC/MS, SEC-MALS, peptide mapping, glycan profiling, cIEF, CHO HCP, residual DNA, ELISA, Western blot, freeze thaw study, micro-developability testing

6.911

50

0

0.5 1 1.5 2 2.5 3 3.5

44.555.566.5 7

7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14 14.5 15

Time (min)

Signal:

DAD1A, Sig=280,4 Ref=o

Name RT (min) 6.911 7.922

Width (min) 0.40 0.33 Sum

Area 23.50

Height 0.79 389.44

Area% 0.28 99.72

Case Study: High-Purity mAb Production Confirmed by SEC-HPLC and Caliper-SDS Data

8365.79 8389.29

In this case, the purity of vanilla antibodies produced in 777 platform showed high purity.

Caliper-SDS

SEC-HPLC

1.dx

Caliper-SDS NR

7.922

100 150 200 250 300 350 400

[Labchip_2021-11-09_18-24-41/A3] 1 NR

99.41

LM % Purity Type

1000 1200

0 200 400 600 800

Xsys

0.59

Xsys

x

6.911

50

0

15

20

25

30

35

40

0.5 1 1.5 2 2.5 3 3.5

44.555.566.5 7

7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14 14.5 15

Aligned Time (sec)

Time (min)

Signal:

DAD1A, Sig=280,4 Ref=o

Caliper-SDS R

Name RT (min) 6.911 7.922

Width (min) 0.40 0.33 Sum

Area 23.50

Height 0.79 389.44

Area% 0.28 99.72

[Labchip_2021-11-09_18-24-41/E3] 1 Reduce

68.06

% Purity Type

8365.79 8389.29

1000 1200 1400 1600

31.94

LM

400 600 800

Xsys

Caliper-SDS

0 200 -200

Caliper-SDS NR

15

20

25

30

35

40

[Labchip_2021-11-09_18-24-41/A3] 1 NR

Aligned Time (sec)

99.41

LM % Purity Type

1000 1200

0 200 400 600 800

Xsys

0.59

Xsys

x

15

20

25

30

35

40

Aligned Time (sec)

Caliper-SDS R

[Labchip_2021-11-09_18-24-41/E3] 1 Reduce

68.06

% Purity Type

1000 1200 1400 1600

31.94

LM

Learn more: Protein Sciences: HTP Production of High-Quality Vanilla Antibodies

About WuXi Biologics

400 600 800

Xsys

WuXi Biologics is a leading contract research, development and manufacturing organization (CRDMO) that provides end-to-end capabilities to healthcare organizations worldwide. With operations in China, the United States, Ireland, Germany, and Singapore, we enable our partners to effectively and efficiently bring biologics and vaccines to patients worldwide through our comprehensive and high-quality drug development model. 15 20 25 30 35 40 Aligned Time (sec) 0 200 -200

The world’s leading global single-source platform from concept to commercialization.

wuxibiologics.com | info@wuxibiologics.com

5-9-2024

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