Application Of Ultrafiltration In The Concentration And Purification Of Avian Influenza Vaccine

Application of Ultrafiltration in the Concentration and Purification of Avian Influenza Vaccine

Avian influenza is an infectious disease of birds caused by influenza A viruses with symptoms ranging from respiratory to severe systemic sepsis. Avian influenza is easily spread among birds and used to be known as "fowl plague", which is classified as a category A infectious disease by the OIE.

Outbreaks of avian influenza occurred in Australia, Italy, Hong Kong, China, and the Netherlands respectively in 1994, 1997, 1999, and 2003, and mainly in Southeast Asia and Europe in 2005. The 2022 avian influenza outbreak reportedly caused poultry producers in 47 U.S. states to lose a total of nearly 59 million poultry, including laying hens, turkeys, and broilers, with government losses exceeding $660 million, making it the deadliest avian influenza outbreak in the history of the U.S. The latest report from October 25, 2023, indicated that cases of HPAI emerged in October in several U.S. states, with the total number of infected poultry exceeding 600,000, of which more than 290,000 had to be culled to prevent the spread of the disease.

PART.1

Avian Influenza Vaccine

The main study of the application of ultrafiltration in the concentration and purification of avian influenza H9N2 vaccine will be presented here.

Avian influenza H9N2 is a low pathogenicity virus subtype belonging to the family Orthomyxoviridae. This subtype is prevalent in the poultry industry as an infectious factor, so there has been a special interest in the development of vaccine production against avian influenza H9N2. Avian influenza H9N2 vaccines are produced by killing the virus and using a mixture of inactivated antigen and oil phase. An egg-based process is currently used to prepare inactivated avian influenza vaccines. The viral antigens of eggs contain large amounts of unwanted proteins that may adversely affect the vaccine formulation. In addition, the production of combined vaccines at low doses requires increased antigen concentrations. This emphasizes the need to increase antigen concentration by improving downstream purification p

rocesses.

After harvesting and clarifying the virus-containing allantoic fluid, it was concentrated and purified with ultrafiltration method. The concentrated and purified samples were assayed for ovalbumin levels.

As can be seen from the SDS-PAGE electropherogram, the concentration and purification can effectively remove the heteroprotein.

Analysis of HI titers in chickens vaccinated with concentrated purified antigen showed that the immune response was above the limiting level (HI=5) for 4 months after vaccination and peaked only on the 99th day after one vaccination. Based on the pattern of immunization duration, antibody levels may be sufficient to cover the limit level within 6 months.

The concentrated and purified vaccine was effective in improving the immune response of vaccinated chicks in the long term. Based on the above results, the use of ultrafiltration technology can be considered to optimize the downstream process (concentration and purification) of influenza (H9N2) vaccine production in order to prepare sustainable vaccines on a large scale without any adverse risk factors.

Guidling Technology's ultrafiltration membrane cassettes have been widely used in the R&D and production of avian influenza epidemics and vaccines, with recovery rates and fluxes close to those of foreign counterparts, and industry teachers are welcome to evaluate.

Guidling Technology is a national high-tech enterprise focusing on biopharmaceuticals, cell culture, purification and concentration of biomedicine, diagnosis and industrial fluids. We have successfully developed centrifugal filter devices, ultrafiltration & microfiltration cassettes, virus filter, TFF system, depth filter, hollow fiber, etc. Which fully meet the application scenarios of biopharmaceuticals, cell culture, and so on. Our membranes and membrane filters are widely used in concentration, extraction and separation of pre-filtration, microfiltration, ultrafiltration and nanofiltration. Our many product lines, from small, single-use laboratory filtration to production filtration systems, sterility testing, fermentation, cell culture and more, meet the needs of testing and production. Guidling Technology is looking forward to cooperating with you!