Production of Inactivated Influenza H5N1 Vaccines from MDCK Cells in Serum-Free Medium

Abstract

BackgroundHighly pathogenic influenza viruses pose a constant threat which could lead to a global pandemic. Vaccination remains the principal measure to reduce morbidity and mortality from such pandemics. The availability and surging demand for pandemic vaccines needs to be addressed in the preparedness plans. This study presents an improved high-yield manufacturing process for the inactivated influenza H5N1 vaccines using Madin-Darby canine kidney (MDCK) cells grown in a serum-free (SF) medium microcarrier cell culture system.Principal FindingThe current study has evaluated the performance of cell adaptation switched from serum-containing (SC) medium to several commercial SF media. The selected SF medium was further evaluated in various bioreactor culture systems for process scale-up evaluation. No significant difference was found in the cell growth in different sizes of bioreactors studied. In the 7.5 L bioreactor runs, the cell concentration reached to 2.3×106 cells/mL after 5 days. The maximum virus titers of 1024 Hemagglutinin (HA) units/50 µL and 7.1±0.3×108 pfu/mL were obtained after 3 days infection. The concentration of HA antigen as determined by SRID was found to be 14.1 µg/mL which was higher than those obtained from the SC medium. A mouse immunogenicity study showed that the formalin-inactivated purified SF vaccine candidate formulated with alum adjuvant could induce protective level of virus neutralization titers similar to those obtained from the SC medium. In addition, the H5N1 viruses produced from either SC or SF media showed the same antigenic reactivity with the NIBRG14 standard antisera.ConclusionsThe advantages of this SF cell-based manufacturing process could reduce the animal serum contamination, the cost and lot-to-lot variation of SC medium production. This study provides useful information to manufacturers that are planning to use SF medium for cell-based influenza vaccine production.

Development of Carbon Nanotubes-Based Immunogens

“…The idea is quite straight forward: the pathogen is obtained in sufficient quantities and then inactivated through the use of different physical and/or chemical methods, including heat, ionizing radiations or chemical agents [8]. The vaccine against seasonal influenza is a good example of an inactivated vaccine in which the Influenza virus is grown on embrionated eggs, purified, and then inactivated by the use of either formaldehyde or beta-propiolactone (currently, an attenuated vaccine against Influenza is also available) [9]. Other examples of inactivated vaccines include immunogens against poliomyelitis (Salk), hepatitis A, rabies and the cellular pertussis vaccine (Pwc-Pertussis whole cell).…”

The Art of Animal Cell Culture for Virus Isolation

“…Vero cells grown in serum-free media has been used for the production of reovirus [39]. MDCK cells grown in serum-free media was used for the production of Influenza H5N1 virus used as a vaccine [40]. Rabies virus used for vaccine production was grown to higher titers in Vero cells in serum free than in serum-containing cell growth media [41].…”

Mycoplasma Removal from Cell Culture Using Antimicrobial Photodynamic Therapy

“…For example, cell culture is used to produce influenza vaccine instead of using eggs. 1 Cell culture has also been used in tissue engineering. Recently, cell sheets derived from healthy donor periodontal ligaments have been reported to regenerate periodontal tissues.…”

“…6 Antimicrobial PDT (aPDT) is applied to destroy bacteria instead of tumor cells. In aPDT, bacteria stained with a dye, which functions as a photosensitizer, 1 should be killed by the ROS induced by light irradiation of the dye. Therefore, aPDT is expected to be a possible alternative to antibiotic treatments for suppressing infections.…”