EB66®: A validated cell line for a more efficient production of viral-based products

EB66® is one of the most extensively-studied and characterized cell lines available for the production of vaccines and other viral-based products1

A hallmark of its success, developments made with EB66® cells have resulted in the approval of a pandemic influenza vaccine for human use, with further approvals in the animal health industry.


Broadly susceptible. EB66® cells have demonstrated the ability to propagate more than 30 different viruses, used either in standard vaccine approaches or in new fields, such as gene therapies or oncolytic viruses.

Manufacturing friendly. EB66® cells are amenable to the modern manufacturing environment: suspension, high-density culture (up to 16 x 107 cells/ml)3, animal component- and serum-free, chemically-defined, GMP-grade media, and a rapid 15-hour doubling time.

Widely adopted. The cell line is in use all over the world for both human and animal health development programs, with approvals in Asia, Europe and South America.

Well characterized. Originating from duck embryonic stem cells, everything from the parental origins of the laying flock to the storage vials of the final cell banks are fully traceable. The cell line has been tested for over 100 different pathogens, including latent and infectious retroviruses, and is free of any contaminant organisms. This information is fully documented in the biologics master file (BMF) submitted to the US FDA in 2008.

Safe and approved by major regulatory authorities. Since the introduction of EB66® cells in 2007, materials derived from the substrate have been administered in various human vaccine clinical trials1,2.

Intellectual property. A vast and extensive patent portfolio provides protection on the cell line, its derivation and use, and may enable further product protection and exclusivity, dependent upon a licensee’s own patent position.

Meeting the challenge for the production of modern viral-based products

An increasing number of human and veterinary pharmaceutical companies are using the EB66® cells for the development of viral-based products in R&D and clinical phases.

In human health

In 2014, a human pandemic influenza vaccine produced in EB66® cells by Kaketsuken (now KM Biologics) received market approval in Japan.

Selected viruses that have been shown to be effectively produced in the EB66® cell line include:

Fields Types (vaccines) Viruses
Infectious disease vaccines Inactivated Japanese encephalitis virus (JEV)
  Live attenuated vaccines Measles, mumps, smallpox, rabies, yellow fever, chikungunya
  Virus like particles Venezuelan equine encephalitis virus (VEEV)
  Sub-Unit vaccines Influenza (commercialized)
  Recombinant vectors Modified virus Ankara (MVA)5, canine distemper virus (CDV), infectious bursal disease (IBDV), fowlpox
  Defective particles Sendai
Oncolytic viruses   Vaccinia, Newcastle disease virus (NDV), measles, herpes simplex virus (HSV), vesicular stomatitis virus (VSV)
Gene therapy   Adeno associated virus (AAV)

Licensees currently using the EB66® cell line include GSK, Sanofi Pasteur, KM Biologics (formerly Kaketsuken), Emergent BioSolutions, Bavarian Nordic, Transgene, Profectus BioSciences, IOZK, and several other undisclosed key players.

In animal health

Three marketing authorizations for EB66®-based veterinary vaccines have already been granted by regulatory authorities worldwide:
  • Vaccine for the prevention of Muscovy duck parvovirus approved by the EMA in 2014
  • Vaccine for the prevention of inclusion body hepatitis virus (Farvet received approval and launched in South America in 2014)
  • Vaccine for the prevention of egg drop syndrome (KM Biologics, formerly Kaketsuken, received marketing authorization in Japan in 2012)

Selected viruses that have been shown to be effectively produced in the EB66® cell line include:

Species Viruses
Avian Influenza, avian reovirus, avian poxvirus (fowlpox, pigeon pox, canary pox), inclusion body hepatitis (IBH), egg drop syndrome (EDS), Newcastle disease virus (NDV), infectious bursal disease (IBDV), duck parvovirus, infectious bronchitis virus (IBV), avian adenovirus, fowl adenovirus, duck adenovirus and enteritis virus
 Canine, Feline Canine distemper (CDV), canine parainfluenza, feline panleukopenia, rabies
 Swine, Equine, Bovine Swine influenza virus, Aujeszky’s disease, Venezuelan equine encephalitis virus (VEEV), bovine parainfluenza, vesicular stomatitis virus (VSV)

Licensees currently using the EB66® cell line include MSD Animal Health, Boehringer Ingelheim Animal Health, Zoetis, Farvet, KM Biologics (formerly Kaketsuken), Fatro, IDT Biologika, and several other undisclosed key players.

Selected EB66® cell line publications
1 A clinical phase I study of an EB66 cell-derived H5N1 pandemic vaccine adjuvanted with AS03. Naruse et al. Vaccine Volume 33, Issue 45, 9 November 2015, Pages 6078-6084
2 Immunogenicity and Safety of an EB66 Cell-Culture-Derived Influenza A/Indonesia/ 5/2005(H5N1) AS03-Adjuvanted Vaccine: A Phase 1 Randomized Trial. Schuind et al. J Infect Dis. 2015 Aug 15; 212(4): 531–541.

3 Process intensification of EB66® cell cultivations leads to high-yield yellow fever and Zika virus production. Nikolay et al. Vaccine. 2015 Nov 9;33(45):6078-84. doi: 10.1016/j.vaccine.2015.09.022. Epub 2015 Sep 25.

4 Culturing a Duck ES-Derived Cell Line in Single-Use Bioreactors: A Rapid, Efficient, and Cost-Effective Vaccine Manufacturing System Based on Suspension Culture. Madeline et al. BioProcess International, Supplement, March 2015, Pages 26 – 33
5 The EB66® cell line as a valuable cell substrate for MVA-based vaccines production, Léon et al. Vaccine Volume 34, Issue 48, 21 November 2016, Pages 5878-5885


For any request or need for additional information please contact:

Wilfried Dang
Manager Business Development

T + 33 228 07 37 10
E bd(at)valneva.com
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