CHO-K1 Stable Cell Line Generation CHO-K1 Stable Cell Line Generation

CHO-K1 Stable Cell Line Generation

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Overview

Cell line generation plays a vital role in the process of drug development. Efficient production of cell lines for recombinant proteins and antibodies that meet industrial requirements can significantly save valuable time and reduce overall costs.

At Biointron, we offer a specialized cell line development service using ECACC-licensed CHO-K1 cells. Our experienced team ensures the rapid creation of high-yield cell lines, tailored to meet our clients' downstream CMC development needs. We pride ourselves on swift and efficient service in the biopharmaceutical industry.

CHO-K1 Stable Cell Line Generation Overview

Highlights

Guaranteed High-Yield

  • Guaranteed high yield of >5 grams/L for antibody production
  • Cell and process optimized
  • Fast turnaround time as short as 3 months

Extensive Experience

  • Proven track record in stable cell line development for more than 10 years
  • Successful delivery of over 100 high-yield cell lines
  • Streamlined and efficient process tailored to your specific needs

Available Commercial Sublicense

  • Research grade or commercial grade available
  • Commercial sublicense available
  • One-time sublicense fee
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CHO-K1 Stable Cell Line Generation
Service Details

Service Step Service Description Timeline Deliverables
Option 1: cell line development
Gene synthesis and transient expression
  • Codon optimization and gene synthesis
  • Subcloning into expression vector
  • Transient expression
 2-3 weeks
  • Gene optimized for the required expression system
  • Gene cloned in expression plasmid
Stable pool generation
  • Stable transfection
  • Stable Pool selection and amplification
 8-9 weeks
Single clone selection and screening
  • Subcloning of the top producing clones
  • Characterization of clones
 8-9 weeks
  • 50mL of the supernatant from stable pool
  • Detailed report
Primary Cell Bank Generation
  • Primary Cell Bank (PCB) preparation
  • PCB Stability Study
 ~3 months
  • Primary cell bank -tested for Mycoplasma
  • Certificate of Analysis
  • Detailed report
CHO-K1 cell report
  • Detail CHO-K1 cell line report and document
 N/A
  • Detail CHO-K1 cell line report and document
Option 2: CHO-K1 cell sublicense service
CHO-K1 sublicense
  • Detail CHO-K1 cell line report and document
  • CHO-K1 sublicense agreement
 3-5 days
  • 2 vial CHO-K1 suspension cell
  • Detail CHO-K1 cell line report and document
  • CHO-K1 sublicense agreement

Case Study

Case 1: Tag free vaccine protein

Single cell image system is applied to confirm the monoclonality. The yield of the final obtained clone A is 2.31 g/L.

Day 0
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Day 1
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Finally

The stability of clone A is also tested by assessing the doubling time of the cells, cell density and yield.

Stability Studies
Subcultured in CD04 medium containing 25 uM MSX the doubling time:22 ± 1hours (Average: 22.6h)
Feed batch
Cells from passages 3, 8, 13, 18, and 23 were collected, and the cell density was assessed. The results indicated a consistent level of cell growth.
Doubling Time of N108-54
Cells from passages 3, 8, 13, 18, and 23 were collected, and the yield was assessed. The results indicated that over 90% productivity titer was retained for over 22 passages.

Case 2: Anti-PD-1

Three single clones were selected, and their yield was assessed using three different commercial media. The results are as follows:

Subclone Medium A (g/L) Medium B (g/L) Medium C (g/L)
414203-N42-6-N1 3.19 5.5 2.63
414203-N42-7-N1 6.48 2.78 1.9
414209-N10-6-N1 7.82 9.02 7.72

Both reducing and non-reducing CE-SDS analyses were performed to assess the purity of these three clones after one step of affinity purification. The results indicated >97% purity.

R-CE-SDS

Sample Name main peak % LMW % Total %
Positive control (Pembrolizumab) 99.1 0.9 100.0
B414203-N42-6-N1 98.8 1.3 100.0
B414203-N42-7-N1 98.5 1.5 100.0
B414209-N10-6-N1 97.8 2.2 100.0

NR-CE-SDS

Sample Name main peak % LMW % HMW % Total %
Positive control (Pembrolizumab) 98.3 1.7 0.0 100.0
B414203-N42-6-N1 94.6 5.4 0.0 100.0
B414203-N42-7-N1 94.1 5.9 0.0 100.0
B414209-N10-6-N1 93.3 6.4 0.3 100.0

CEX-HPLC analyses were performed to assess the charge of these three clones. The results indicated a similar main component% compared to the positive control for the first two clones. Additionally, the first clone (B414203-N42-6-N1) shows the most similar basic component%.

CEX-HPLC

Sample Name Acidic component % Main component % Basic component % Total %
Positive control (Pembrolizumab) 19.5 64.1 16.4 100.0
B414203-N42-6-N1 16.0 69.7 14.3 100.0
B414203-N42-7-N1 17.4 69.2 13.4 100.0
B414209-N10-6-N1 18.1 44.6 37.3 100.0

Currently, we have sublicensed the CHOK1BN cell line to dozens of customers, and the status of some customers' projects is as follows:

Customer Type CLD PD Pilot Nonclinical IND Phase I Phase II
1 Customer A ADC
2 Customer G R-vaccines
3 Customer H Mab
4 Customer B Mab
5 Customer C Fab
6 Customer J R-vaccines
7 Customer F R-glycoprotein
“With ECACC-licensed CHO-K1 cells, we can generate research grade or commercial grade stable cell lines, with the option for a one-time sublicensing fee for a perpetual and irrevocable license.”
Jing Cheng
Jing Cheng
Cell Line Expression Team

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