Column chromatography for the development of pandemic and seasonal influenza vaccines Olga Chervyakova Research Institute for Biological Safety Problems, RK ME&S 5 th Meeting with International Partners on Prospects for Influenza Vaccine Technology Transfer to Developing Country Vaccine Manufacturers Belgrade, Serbia, 27-28 March 2012
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Column chromatography for the development of pandemic and … · Column chromatography for the development of pandemic and seasonal influenza vaccines Olga Chervyakova Research Institute
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Column chromatography for the development of pandemic and seasonal influenza vaccines
Olga ChervyakovaResearch Institute for Biological Safety Problems, RK ME&S5th Meeting with International Partners on Prospects for Influenza Vaccine Technology Transfer to Developing Country Vaccine Manufacturers
Protein removal (%) 90.20 96.62 97.28 99.19 97.55 94.88
HA total (SRID) (%) 100 100 100 100 100 100
Gel-filtration (Sepharose CL-6B)
Protein removal (%) 96.27 99.35 98.58 99.50 98.07 96.22
HA total (SRID) (%) 59.0 46.0 51.4 52.1 100.0 85.7
Inactivation
Detergent treatment/removal
0.22 µm filtration
HA total (SRID) (%) 14.8 17.0 33.0 37.4 60.4 32.9
Ovalbumin (µg/mL) 0.75 1.5 0.65 0.25 1.2 0.5
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Bulk Specifications : H1N1
Seasonal influenza vaccines: Development and Research
Items Qualifying standardsBatch Results
2012/1 2012/2 2012/3 2012/4
Identity wt-like Confirm Confirm Confirm Confirm
Total Protein(µg/ml)
≤ 6-foldHA content
0.475 0.325 0.490 0.247
HA Content(µg/ml)
≥ 90 113.82 76.12 124.46 47.09
Endotoxin(EU/ml)
≤ 100 5.0 1.25 1.25 1.25
Ovalbumin(µg/ml)
≤ 2 1.3 0.25 1.2 0.5
Sterility To meet requirement Confirm Confirm Confirm Confirm
16Seasonal influenza vaccines: Development and Research
Conclusion
•Tangential diafiltration in combination with column chromatography may be successfully used for influenza virus purification.
•The method is suitable for different influenza virus subtypes. In dialysis and chromatography the same buffers are used regardless of the virus subtype.
•Chromatographic profiles of the different influenza virus subtypes are identical. Virus is eluted in the void volume.
•≥99.0% removal of protein; ovalbumin content <2 µg/mL; HA yields > 30%