METHODS OF THE PROTEINS DOWNSTREAM PURIFICATION EFFICIENCY INCREASING WITH THE USE OF MEMBRANE ION EXCHANGE CHROMATOGRAPHY

摘要

FIELD: technological processes; chemistry.;SUBSTANCE: invention relates to the downstream antibodies purifying chromatography steps efficiency enhancing method, comprising: a) passing a composition comprising an antibody of interest and a contaminating ionic polymer through an ion-exchange membrane, characterized by that the antibody and membrane have an opposite charge, under operating conditions, which consist of a buffer having a pH sufficiently different from the antibody pI to increase the antibody charge, and a low ionic strength effective to prevent buffer ion charges shielding, which cause the membrane to bind the antibody and at least one contaminating ionic polymer; b) ion exchange membrane reloading to a loading density of 1,000–5,000 g/L in such a way, that at least one of said contaminating ionic polymer remains bound to the membrane, whereas the antibody of interest is mainly in the eluate; c) trapping the eluate from the ion exchange membrane, which contains the antibody of interest; d) ion exchange chromatography step, which has a charge similar to the previous membrane, to which the membrane eluate containing the antibody of interest is subjected, and e) recovering the purified antibody from the ion exchange chromatography eluate.;EFFECT: method makes it possible to improve the proteins purification and to remove the ionic polymer impurities, for example, such as polyethyleneimine.;27 cl, 16 dwg, 2 tbl, 1 ex