菜单
Service Overviews

Background: An IND application project needs to complete the downstream purification process development and prepare proteins with acceptable quality.


Methods: Purification process development is performed by applying design of experiments (DoE), combining with the customer-oriented resins and excipients of high quality and purity, to meet the compliance requirement.

1. The cell culture materials are used for resin screening study along in the pool and mAb screening stages.

2. With well-defined monoclonal antibody, purification process is optimized and validated for the qualification .

3. Upon on QbD based process optimization, DoE is preferentially applied to the purification process development.

4. The specifications of materials, reagents, and excipients in use are strictly selected and reviewed to meet the regulatory compliance requirement.

Service Overviews Background: An IND application project needs to complete the downstream purification process development and prepare proteins with acceptable quality. Methods: Purification process development is performed by applying design of experiments (DoE), combining with the customer-oriented resins and excipients of high quality and purity, to meet the compliance requirement. 1. The cell culture materials are used for resin screening study along in the pool and mAb screening stages. 2. With well-defined monoclonal antibody, purification process is optimized and validated for the qualification . 3. Upon on QbD based process optimization, DoE is preferentially applied to the purification process development. 4. The specifications of materials, reagents, and excipients in use are strictly selected and reviewed to meet the regulatory compliance requirement. Service Contents 1. Development and qualification of the purification process for the PCC(DNA)-IND project as a whole. 2. Gram-level protein preparation for phased projects. 3. Validation of endotoxin and other impurities removals or virus clearance for phased projects. Service Highlights 1. High Efficiency and High Yield With the complete and fully functioned facilities, adequate talent of manpower management, and protein stability study coordinated with purification process development, the overall process development cycle < 2 months. The loading pH, conductivity, protein concentration, retention time, and other parameters are optimized to increase capacity, and the elution pH, conductivity, buffer type, and other conditions are optimized to improve the yield of target protein. The yields of depth filtration, affinity chromatography, cation exchange chromatography, anion exchange chromatography, nanofiltration, and ultrafiltration are more than 95%, 95%, 85%, 95%, 98%, and 95%, respectively; the overall recovery rate is more than 60%. 2. High Quality Protein quality: The purity of SEC-HPLC is more than 97%, the purity of non-reduced CE-SDS is more than 95%, and the CEX main peak is more than 50%. All tests meet the requirements of Chinese Pharmacopoeia and USP. Residual impurities such as HCP, HCD, rProteinA, endotoxin, and bioburden satisfied the requirements of Chinese Pharmacopoeia and USP. The Quality such as post-translational modification satisfied the requirements of Chinese Pharmacopoeia and USP. 3. Customer-Oriented Under the premise of ensuring compliance of resins and excipients, the sources designated or suggested by the customer were preferred. Customized study protocols are provided to customers. A mature modular platform, advanced testing instruments and equipment, and simultaneous investigation of multiple test indicators boost the early application of projects. Case Stastics 1. Affinity chromatography for a bispecific antibody project Background: CMC manufacturing process development was required for a bispecific antibody in IND for the treatment of pancreatic cancer. In this case, the affinity chromatography process step of the purification process was developed. Objective: To perform resin screening and capacity test, optimize the impurity removal conditions, and improve yield. Method: 10% antibody titer flow-through method Fig. 1 Affinity chromatogram of a bispecific antibody project 2. Cation exchange chromatography for a bispecific antibody project Background: CMC manufacturing process development was required for a bispecific antibody in IND for the treatment of pancreatic cancer. In this case, the cation exchange chromatography process step of the purification process was developed. Objectives: To perform resin screening and capacity test, optimize the impurity removal conditions, and improve yield. Methods: Based on the concept of QbD, full factorial DoE was adopted, the process parameters of cation exchange chromatography were optimized to obtain the process operation ranges for the highest capacity and the lowest residual content of HCP, HCD, and other impurities, to improve the target protein quality such as SEC, CE, and CEX. Results: The capacity reached 55 mg/mL with the yield of over 85%; the residual impurities were acceptable. Conclusions: The cation chromatography process was successfully developed, and the process performance and parameters met appropriate requirements. Fig. 2 Cation exchange chromatogram of a bispecific antibody project 3. Anion exchange chromatography of a bispecific antibody project Background: CMC manufacturing process development was required for a bispecific antibody in IND for the treatment of pancreatic cancer. In this case, the anion exchange chromatography process step of the purification process was developed. Objectives: To optimize resin screening and the conditions for removal of impurities such as HCP, HCD, rProteinA, and endotoxin. Methods: Based on the concept of QbD, flow-through mode and full factorial DoE were adopted, and the process parameters of anion exchange chromatography were optimized to obtain the process operation ranges for high capacity and low residual content of HCP, HCD, and other impurities. The optimized process can effectively remove related viruses and ensure the biosafety of final products. Results: The capacity reached 150 mg/mL with the yield of over 95%; the residual impurities were acceptable. Conclusions: The anion chromatography process was successfully developed, and the process performance and parameters satisfied appropriate requirements. Fig. 3 Anion exchange chromatogram of a bispecific antibody project 4. Overall downstream purification process indicators of a bispecific antibody project Background: CMC manufacturing process development was required for a bispecific antibody in IND for the treatment of pancreatic cancer. In this case, the overall process performance was summarized. Objectives: To summarize process parameters and assess process performance. Methods: The process performance and residual impurities were summarized in a stepwise manner. Results: The process performance parameters satisfied appropriate production requirements, and the residual impurities satisfied appropriate regulatory requirements. Conclusions: The process was successfully developed and can be employed for the production of protein samples for IND application. Table 1 Purification process step indicators of a bispecific antibody project Flow Capacity/parameter Recovery SEC NR-CE CEX HCP (ppm) HCDNA (pg/dose) rProteinA (ppm) Depth filtration ≥ 150 L/m2 ≥ 95% / / / ≥ 50000 / / Affinity chromatography ≤ 48 mg/mL ≥ 93% ≥ 95% ≥ 94% ≥ 60% ≤ 5000 ≤ 200 ≤ 100 Virus inactivation by low pH incubation pH 3.5–3.7 ≥ 95% ≥ 95% ≥ 94% ≥ 60% / / / Intermediate depth filtration ≤ 1000 g/m2 ≥ 95% ≥ 95% ≥ 94% ≥ 60% 600–1000 ≤ 100 ≤ 80 Cation exchange chromatography ≤ 55 mg/mL ≥ 85% > 98% ≥ 95% ≥ 65% 100–200 ≤ 50 ≤ 30 Anion exchange chromatography ≤ 150 mg/mL ≥ 95% > 98% ≥ 95% ≥ 65% ≤ 20 ≤ 20 ≤ 10 Nanofiltration ≥ 1500 g/m2 ≥ 98% > 98% ≥ 95% ≥ 65% ≤ 20 ≤ 20 ≤ 10 Ultrafiltration ≤ 1500 g/m2 ≥ 95% > 98% ≥ 95% ≥ 65% ≤ 20 ≤ 20 ≤ 10

Service Contents

1. Development and qualification of the purification process for the PCC(DNA)-IND project as a whole.

2. Gram-level protein preparation for phased projects.

3. Validation of endotoxin and other impurities removals or virus clearance for phased projects.


Service Highlights
  • 1. High Efficiency and High Yield
    1. With the complete and fully functioned facilities, adequate talent of manpower management, and protein stability study coordinated with purification process development, the overall process development cycle < 2 months.
    2. The loading pH, conductivity, protein concentration, retention time, and other parameters are optimized to increase capacity, and the elution pH, conductivity, buffer type, and other conditions are optimized to improve the yield of target protein.
    3. The yields of depth filtration, affinity chromatography, cation exchange chromatography, anion exchange chromatography, nanofiltration, and ultrafiltration are more than 95%, 95%, 85%, 95%, 98%, and 95%, respectively; the overall recovery rate is more than 60%.
  • 2. High Quality
    1. Protein quality: The purity of SEC-HPLC is more than 97%, the purity of non-reduced CE-SDS is more than 95%, and the CEX main peak is more than 50%. All tests meet the requirements of Chinese Pharmacopoeia and USP.
    2. Residual impurities such as HCP, HCD, rProteinA, endotoxin, and bioburden satisfied the requirements of Chinese Pharmacopoeia and USP.
    3. The Quality such as post-translational modification satisfied the requirements of Chinese Pharmacopoeia and USP.
  • 3. Customer-Oriented
    1. Under the premise of ensuring compliance of resins and excipients, the sources designated or suggested by the customer were preferred.
    2. Customized study protocols are provided to customers.
    3. A mature modular platform, advanced testing instruments and equipment, and simultaneous investigation of multiple test indicators boost the early application of projects.

Case Studies
1. Affinity chromatography for a bispecific antibody project

Background: CMC manufacturing process development was required for a bispecific antibody in IND for the treatment of pancreatic cancer. In this case, the affinity chromatography process step of the purification process was developed.

Objective: To perform resin screening and capacity test, optimize the impurity removal conditions, and improve yield.

Method: 10% antibody titer flow-through method

Fig. 1 Affinity chromatogram of a bispecific antibody project

2. Cation exchange chromatography for a bispecific antibody project

Background: CMC manufacturing process development was required for a bispecific antibody in IND for the treatment of pancreatic cancer. In this case, the cation exchange chromatography process step of the purification process was developed. 

Objectives: To perform resin screening and capacity test, optimize the impurity removal conditions, and improve yield.

Methods: Based on the concept of QbD, full factorial DoE was adopted, the process parameters of cation exchange chromatography were optimized to obtain the process operation ranges for the highest capacity and the lowest residual content of HCP, HCD, and other impurities, to improve the target protein quality such as SEC, CE, and CEX. 

Results: The capacity reached 55 mg/mL with the yield of over 85%; the residual impurities were acceptable.

Conclusions: The cation chromatography process was successfully developed, and the process performance and parameters met appropriate requirements.


Fig. 2 Cation exchange chromatogram of a bispecific antibody project

3. Anion exchange chromatography of a bispecific antibody project

Background: CMC manufacturing process development was required for a bispecific antibody in IND for the treatment of pancreatic cancer. In this case, the anion exchange chromatography process step of the purification process was developed. 

Objectives: To optimize resin screening and the conditions for removal of impurities such as HCP, HCD, rProteinA, and endotoxin. 

Methods: Based on the concept of QbD, flow-through mode and full factorial DoE were adopted, and the process parameters of anion exchange chromatography were optimized to obtain the process operation ranges for high capacity and low residual content of HCP, HCD, and other impurities. The optimized process can effectively remove related viruses and ensure the biosafety of final products. 

Results: The capacity reached 150 mg/mL with the yield of over 95%; the residual impurities were acceptable. 

Conclusions: The anion chromatography process was successfully developed, and the process performance and parameters satisfied appropriate requirements.


Fig. 3 Anion exchange chromatogram of a bispecific antibody project

4. Overall downstream purification process indicators of a bispecific antibody project

Background: CMC manufacturing process development was required for a bispecific antibody in IND for the treatment of pancreatic cancer. In this case, the overall process performance was summarized. 

Objectives: To summarize process parameters and assess process performance. 

Methods: The process performance and residual impurities were summarized in a stepwise manner. 

Results: The process performance parameters satisfied appropriate production requirements, and the residual impurities satisfied appropriate regulatory requirements. 

Conclusions: The process was successfully developed and can be employed for the production of protein samples for IND application.


Table 1 Purification process step indicators of a bispecific antibody project

Flow

Capacity/parameter

Recovery

SEC

NR-CE

CEX

HCP

(ppm)

HCDNA

(pg/dose)

rProteinA (ppm)

Depth filtration

≥ 150 L/m²

≥ 95%

/

/

/

≥ 50000

/

/

Affinity chromatography

≤ 48 mg/mL

≥ 93%

≥ 95%

≥ 94%

≥ 60%

≤ 5000

≤ 200

≤ 100

Virus inactivation by low pH incubation

pH 3.5–3.7

≥ 95%

≥ 95%

≥ 94%

≥ 60%

/

/

/

Intermediate depth filtration

≤ 1000 g/m²

≥ 95%

≥ 95%

≥ 94%

≥ 60%

600–1000

≤ 100

≤ 80

Cation exchange chromatography

≤ 55 mg/mL

≥ 85%

> 98%

≥ 95%

≥ 65%

100–200

≤ 50

≤ 30

Anion exchange chromatography

≤ 150 mg/mL


≥ 95%

> 98%

≥ 95%

≥ 65%


≤ 20

≤ 20

≤ 10

Nanofiltration

≥ 1500 g/m²

≥ 98%

> 98%

≥ 95%

≥ 65%

≤ 20

≤ 20

≤ 10

Ultrafiltration

≤ 1500 g/m²

≥ 95%

> 98%

≥ 95%

≥ 65%

≤ 20

≤ 20

≤ 10