We work with most cell lines used for manufacturing monoclonal antibodies and recombinant proteins, including CHO, NS/0, BHK, 293 as well as murine hybridomas, and all selection systems used with these lines.
We have also worked with several different selection systems such as dhfr / methotrexate CHO, mycophenolic acid/xanthine, and G418 systems. Our experience includes attached or surface dependent as well as suspension culture types.
GBI offers a full range of mammalian cell culture development and manufacturing services ranging from the creation of up and downstream processes sufficient for the manufacturing of proof of concept or toxicology product material to processes for licensed manufacturing. We do that with one goal in mind—to provide superior service to our clients that not only meets, but exceeds their expectations.
We work with most cell lines used for manufacturing, including CHO, NS/0, BHK, 293 as well as murine hybridomas, and all selection systems used with these lines. We have also worked with several different selection systems such as dhfr - /methotrexate CHO, mycophenolic acid/xanthine, and G418 systems. Our experience includes surface dependent as well as suspension culture types.
Our clients have come from Eastern and Western Europe, North America and Asia, and have included biotechnology startup firms, research institutes, university laboratories, multinational biotechnology and pharmaceutical companies, and government institutes with projects that range from Translational Medicine and Proof of Concept projects to producing material for clinical trials.
Cell Line Development
GBI scientists have considerable experience in the development of cell lines for cGMP manufacturing for clinical studies through commercial manufacturing. In addition, working with our partners, we can develop cell lines from cDNA that you provide. Our partners offer their own proprietary vectors to develop cell lines that can routinely produce up to 4g/L in CHO cell lines.
Our Approach to Cell Line Development
GBI will work with one of our collaborative partners for molecular biology and cell line development activities.
For each project, we will prepare a cell line development project plan (design input) for review and approval by the client.
Cell line development can either begin with an existing vector as supplied by the client, or vector design and construction (gene synthesis) can be performed.
Through our collaborative partnerships, we can take your gene sequence or cDNA for your recombinant protein, antibody, or hybridoma and engineer a highly productive, stable cell line in CHO, BHK, NS0, HEK-293, SP2/0 cells or proprietary third-party systems (GS, ACE, UCOE, PER.C6®, etc.).
After transfection of the cells, screening amplification and subcloning will be performed. Cell lines will be adapted to suspension culture in serum-free, chemically-defined media. Also, note that per client request, stable pool of transfectants can be generated and characterized prior to the generation of a high producing single stable clone.
Typically, cell line development activities will result in the identification of three (3) to five (5) high-producing candidate clones and ten (10) liquid nitrogen vials of each of three (3) to five (5) clones will be created.
During cell line development at the shake flask stage, a small-scale, one (1) column protein A purification of the harvest can be performed for each cell line for initial characterization of the protein structural (primary, secondary, tertiary and quaternary) and binding characteristics via methods such as mass spectrometry, binding kinetics (ForteBio Blitz or Octet systems), glycosylation, potency, amino acid analysis, etc., to better aid in selection of the optimal cell line along with subsequent generational stability and productivity data.
For more information on our cell line development capabilities, please click here.
Generational Stability Studies of Cell Line and RCB Creation
For each project, we will prepare a project plan for review and approval by the client.
The stability of the three (3) to five (5) candidate cell lines will be evaluated in shake flask culture for up to fifty (50) generations, for example. Cell growth kinetics and productivity will be evaluated at regular intervals, e.g., every five (5) generations.
Together with our client we will evaluate the data and choose the best performing clone for creation of Research Cell Bank (RCB).
At least forty (40) vials of the RCB will be produced.
The RCB will be tested for mycoplasma, sterility, and adventitious viruses, and then shipped to GBI for process development work and creation of a GMP MCB.
A final report for cell line development will be prepared for review and approval by the client for each project.
To learn more on our approach to cell line development, click here.
GBI offers a full array cell banking services for research or manufacturing use. Manufacturing banks include seed, master, working, and post production. All of these banks are GMP compliant and are tested based on the current Points to Consider for cell bank testing. GBI works closely with our clients and with our qualified contract testing organizations to assure acceptability of all testing. Banks are prepared using a controlled freeze rate system and are stored in -150°C mechanical freezer systems. All bank inventories are secured and controlled by GBI's Quality department. Short- or long-term storage of cell banks is provided.
Research Cell Bank
Research cell banks can be made at GBI to carry out non-GMP process development activities.
GMP (Master and Working)
GBI produces Master Cell Banks (MCBs) and Working Cell Banks (WCBs) in compliance with cGMP requirements.
Cell Culture Process Development
GBI will develop a robust and reproducible cell culture process for the production of complex therapeutic protein drug candidates. GBI has experience with a number of cell cultivation technologies including stationary culture disposable flasks, spinners, WAVE disposable and stirred tank bioreactors. GBI offers cell culture production services for non-bioreactor projects.
Media selection and development (both animal component and animal-free components)
GBI offers medium evaluation and adaptation to commercial serum free formulations for your cell line. Our goal is to increase your line's productivity, growth performance, and/or compliance level.
Development and optimization of robust and scalable processes
GBI offers three different types of bioreactor systems for production and process development services for each. We work with clients to determine the optimum bioreactor type to use for their project based on their cell expression system and product quantities needed.
Stirred tank batch or fed-batch bioreactor
Stirred tank bioreactors up to 500L are available. With the possibility of achieving yields up to three grams per liter, our bioreactors could produce up to 1,500 grams per every 8 to 10 day run.
Fibracell disk immobilized bed perfusion bioreactors
Bioreactors with cartridge surface areas ranging from 2.1 to 25.2m² are available. The larger of these bioreactors can produce in excess of 150 grams during a six-week production run.
6 X 5L Sartorius Bioreactors
GBI has available small bioreactor systems of each type for the development of specific run parameters and scaling data for the larger production bioreactors. These small systems are also available for the production of non-GMP material for testing and toxicology studies.
GBI manufactures GMP material for clients using:
Dakota stirred tanks, 200L and 500L.
50L disposable WAVE bioreactor
The scale of bioreactor selected depends on the productivity of the cell line and the requirement for GMP material. Once the GMP material is produced, GBI performs in-process and release testing as well as QA review of the GMP manufacturing documentation. In addition, GBI offers stability studies, viral clearance studies, creation of a reference standard, etc. to meet regulatory compliance needs.
Anthony Attipoe Manager of Cell Culture Manufacturing
Anthony Attipoe has over 25 years of experience
with major biotechnology companies such as Genentech, Cephalon, Collaborative BioAlliance, Baxter HT, Nabi Biopharmaceuticals, and Dendrion. His expertise in fermentation and cell culture extends through all phases of process development to clinical trials and commercial production of recombinant proteins and vaccines.
Recently, he has focused on mammalian cell culture development, scale-up, tech transfer, large- and small-scale bioreactor operations, and optimization of processes that can lead to significant increases in biopharmaceutical product yield. He has also successfully managed and trained bioprocess teams in quality systems and compliance programs.
Anthony earned his Bachelor’s degree in Biology from the State University of New York at Binghampton.
Click here to learn more…
Donggou He, MD, PhD Manager of Cell Culture Development
Dr. Donggou “Diego” He comes to GBI with
more than 15 years in academia focusing on clinical immunology, which is augmented by several years of industrial experience dedicated to Cell Culture Process Development and Molecular Biology where he lead CHO-based cell culture process development and process optimization by design of experiment (DOE) for innovative proteins as well as Biosimilars, which led to a high yield of up to 5g/L. He has designed and conducted cell-based assays and immunochemical assays to support the discovery and development of therapeutic recombinant monoclonal antibodies. Diego received his Medical Degree in Clinical Medicine from the South China University Medical School and subsequently received his PhD in Molecular Biology from Sun Yat-Sen University.