MANUFACTURING METHOD Russian patent published in 2022 - IPC C07K14/47 C07K1/14 C07K1/18 C07K1/22 A61K38/17 

FIELD: biotechnology.

SUBSTANCE: group of inventions relates to biotechnology, in particular to methods for isolation and purification of recombinantly expressed intracellular protein annexin A5 (hereinafter – AnxA5) from a host cell producing endotoxin with a cellular wall, and to its use. Methods include release of intracellular protein AnxA5 from the host cell in the presence of buffer for homogenization containing non-ionic detergent selected from Twin 20 and Twin 80. Then, a composition containing protein AnxA5 and chelator of metal calcium ions AnxA5 is exposed to anion-exchange resin in the presence of additional selected metal ions. At the same time, protein AnxA5 remains in the solution at all stages of methods except for cases, when it temporarily binds to any chromatographic resins.

EFFECT: inventions exclude the need for use of any stages of purifying centrifugation for protein AnxA5 after isolation from a host cell, allow for minimization of a degree of binding endotoxin and AnxA5 at initial stages of isolation, and contribute to the production of a purified product of annexin A5 on an industrial scale, suitable for therapeutic use.

98 cl, 8 dwg, 11 tbl, 4 ex