Development of H7N9 Influenza Vaccine

The H7N9 candidate vaccine strains virus (CCVs) (NIBRG-268) was obtained from the NIBSC. This virus contains six internal genes from the egg-adapted high-growth A/PR8 virus and two surface protein genes (HA and NA) from A/Anhui/1/2013 (H7N9). This strain was further adapted to MDCK cells (ATCC CCL-34), which were purchased from the Food Industry Research and Development Institute, Hsinchu, Taiwan. The influenza strain H7N9 were originally derived from the egg process. The egg-derived CCVs do not grow well in MDCK cell culture. The re-adapted process was done by choosing better replication colonies in the plaque assay. Generally, the process needs to be repeated 3 to 10 times before obtaining the high growth CVVs in the MDCK cell culture. For preparation of the vaccine standard antigen, OptiPro serum-free medium (Invitrogen) was used for MDCK cell growth, and OptiPro supplemented with 2 μg/mL TPCK-trypsin (Sigma) was used for viral replication.^{12 (link)} The harvested virus was inactivated with formaldehyde and then purified by clarification and chromatography.^{13 (link)} This final bulk-produced H7N9 virus was considered the cell-derived PLS.

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Lai C.C., Weng T.C., Chen P.L., Tseng Y.F., Lin C.Y., Chia M.Y., Sung W.C., Lee M.S, & Hu A.Y. (2020). Development and characterization of standard reagents for cell-based prepandemic influenza vaccine products. Human Vaccines & Immunotherapeutics, 16(9), 2245-2251.

Publication 2020

OptiPro TPCK-trypsin

Antigen Assay Bulk Ccl 34 Cell Cell culture Chromatography Formaldehyde H7n9 H7n9 virus Influenza Mdck cell Plaque Protein genes Replication Serum Standard preparation Strains Tpck Trypsin 2 Vaccine Vaccine virus Viral replication Virus Virus genes

Corresponding Organization : National Health Research Institutes

Other organizations : National Tsing Hua University, National Chung Hsing University

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Variable analysis

independent variables

Re-adaptation process of the H7N9 candidate vaccine strains virus (CCVs) (NIBRG-268) to MDCK cells

dependent variables

Growth of the re-adapted H7N9 CCVs in MDCK cell culture

control variables

Use of OptiPro serum-free medium for MDCK cell growth
Use of OptiPro medium supplemented with 2 μg/mL TPCK-trypsin for viral replication
Inactivation of the harvested virus with formaldehyde
Purification of the virus by clarification and chromatography

controls

No positive or negative controls were explicitly mentioned in the information provided.

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