Service Overview
Background: Industrial cell lines are the foundation of commercial production of antibody drugs. Compliance, expression level, product quality and stability are 4 key indicators to evaluate the cell lines. Currently, the construction of high-expression and stable industrial cell lines in the industry generally meets problems such as unclear traceability of host cells, long cell line construction duration, unguaranteed monoclonal origin and low antibody expression level.
Method: Sanyou Bio launched the high-expression and stable industrial cell line construction service, aiming to provide global clients with fast, efficient, compliant, high expression, high quality and stable industrial cell lines. Based on client requirements, our service can cover gene optimization, vector development, cell transfection, cell pool screening, single cell cloning and screening.
Advantages: The platform has licensed in 4 sets of commercially authorized cell lines and implemented cGMP standards. It takes only 2.5 months to obtain stable, high-expression, and high-quality cell lines with the expression level as high as 12 g/L.
Cases: Our team members have extensive project experience in the construction and identification of dozens of industrial cell lines, including production cell lines for bispecific antibodies, trispecific antibodies, monoclonal antibodies and fusion proteins.
Service Highlights
High titer： The expression levels up to 12 g/L
The production is based on multiple screening systems, such as plate bioreactors, tubular bioreactors, shake flasks, and fermenters. We adopt GS high-expression screeningsystem and provide 4 types of commercially authorized host cells. Bulk pool and mini pool construction modes are used and over 1000 clones are screened to obtain stable high-expression industrial cell lines.
Stable production：Over 60 PDL assessments are performed to support the commercialized production.
Over 60 PDL assessments for cell line passage stability, cellular growth stability, antibodycontinuous production stability, genetic sequence consistency, and gene copy number are conducted to support the commercialized production.
Premium quality：Comprehensive quality control and stability evaluations are performed with the drug products.
11 product quality control indicators and 10 cell line stability indicators are included. Clone screening is guided by the comprehensive test results for high-quality cell lines.
Compliance： The seed cells conform to cGMP standards and 4 sets of host cells are commercially authorized.
All file records including the host cell traceability, cell line passage records, cell bank construction and verification records are clear and traceable. The monoclonality of cell lines meets the requirements of regulations and the image data are intact. The construction of cell bank conforms to cGMP standards. The verification of cell bank meets the regulatory requirements of China and the United States.
Fast speed：Only 2.5 months to obtain stable and productive monoclonal cell lines.
Stable and productive industrial monoclonal cell lines for therapeutic product development can be obtained in approximately 2.5 months.
Service Content
Process
Case Study
1. Cell line expression data
1.1 Data summary of cell line construction stage of a monoclonal antibody project
Objective: To investigate the increasing trend of cell expression level during cell line construction.
Methods: The expression level data of cell pool construction and monoclonal cell line construction at various stages of a monoclonal antibody development project were summarized.
Conclusion: With the same feeding and fermentation conditions, the expression level of monoclonal cell lines increased by 2.4 times compared with that of the cell pools. After preliminary optimization, the expression level of monoclonal cell lines in shake flask reached 9.0 g/L.
Fig. 1 Data summary of various cell line development stages
1.2 Expression data of monoclonal cell lines in shake flasks
Objective: To investigate the expression level of cell line construction platform.
Methods: The expression levels of the top four monoclonal cell lines of 24 projects with 14-day feeding and fermentation in shake flasks were analyzed and the results are shown in fig. 2.
Conclusion: The expression levels of monoclonal cell lines are between 4 g/L-6 g/L with the highest level reaching 10.41 g/L, indicating that the platform process has been well-established.
Fig. 2 Expression level of monoclonal cell lines fed-batch culture in shake flasks
1.3 Expression level and Qp of monoclonal cell lines in 24 deep-well plates
Objective: To investigate the expression level and Qp of monoclonal cell lines in 24 deep-well plates.
Methods: Twenty-five selected monoclonal cell lines were put into 24-deep-well plates for 12 days of feeding and fermentation. Samples were collected regularly for cell density and viability assay. At the end of culture, antibody expression level was detected and Qp values were calculated according to the integral values of expression level and viable cell density.
Conclusion: The screening results are shown in fig. 3. Eleven cell lines in 24-deep-well plates had expression level of over 2.5 g/L with the highest of 3.9 g/L and the Qp reached 57 pg/cell/day.
Fig. 3 Expression level and Qp scattering of cell lines fed-batch culture in 24-deep-well plates
2. Data from a bispecific antibody development project
2.1 3L bioreactor feeding and fermentation data
Objective: To investigate the cell growth and expression of monoclonal cell lines in the bioreactor.
Methods: Cell lines were seeded into 3L bioreactors for feeding and fermentation and samples were collected to test relevant indicators.
Conclusion: Viable cell density (VCD) reached peak value ((1.0-1.5)E+7 cells/mL) on day 6 to day 7 and the concentration of metabolic byproduct (lactic acid) maintained at ＜ 2 g/L. After culture of 16 days, the cell viability remained 70% and the antibody expression level reached 12 g/L.
Fig. 4 3L bioreactor culture data from a bispecific antibody development project
2.2 Summary of cell line stability data of a bispecific antibody development project
Objective: To investigate the stability of monoclonal cell lines.
Methods: In a bispecific antibody development project, cell bank stability study was investigated after development of the GMP cell bank.
Conclusion: The deviation of gene copy numbers was within 30% and when PDL exceeded 60, the deviation of expression level was within 30%, which met the industry requirements. The cell line is stable enough to support the production at 50, 000 L scale.
Fig. 5 Stability evaluation (A. expression level; B. gene copy number)
2.3 Quality analysis data
2.3.1 SEC-HPLC
In a bispecific antibody development project, SEC-HPLC was performed on expression products in the shake flask screening stage of cell line development. The results show that the main peak ratio of this bispecific antibody sample is 96.57%.
Fig. 6 SEC-HPLC
2.3.2 CE-SDS
In a bispecific antibody development project, CE-SDS was performed on expression products in the shake flask screening stage of cell line development. The results show that the main peak ratio of this bispecific antibody sample is 95.4%.
Fig. 7 CE-SDS
2.3.3 CEX
In a bispecific antibody development project, CEX was performed on expression products in the shake flask screening stage of cell line development. The results show that the main peak ratio of this bispecific antibody sample is 65.56%.
Fig. 8 CEX
Facilities
(1) C + A class GMP clean environment occupying around 1000 m^2
(2) Nearly a hundred sets of cutting-edge instruments and equipment
1000+ m^2 C + A class clean environment
Cell shaking incubator
Single cell printer
Monoclonal imager
Maurice protein characterization analysis system
High-throughput surface plasmon resonance system
Achievements
Inquriy
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