FIELD: biotechnology.
SUBSTANCE: method for production of recombinant pseudoadenovirus particles (RPAN) concentrate, expressing the hemagglutinin gene of influenza a/california/07/2009(H1N1) is proposed. The method comprises: obtaining of a production cell culture; preparation of disaggregated starting RPAN culture samples; single-round infection of the production suspension cell culture, thereby obtaining a RPAN-containing crude suspension, with subsequent cell mass precipitation by centrifugation and spent culture medium diverting; precipitated cell mass resuspension in lysis buffer, then one-time freezing followed by suspension thawing the slurry and RPAN separation from the disrupted cells successively by centrifugation, ultrafiltration, anion exchange chromatography, size exclusion chromatography, sterile filtration to yield a sterile pharmaceutical grade RPAN concentrate with high level ofchromatographic purity.
EFFECT: method allows to obtain an increased yield of RPAN and a possibility to obtain a concentrate with high titers and chromatographic purity, that meets the quality of a pharmaceutical product suitable for industrial production of influenza vaccines.
1 dwg, 1 tbl, 5 ex
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