METHODS FOR PRODUCING DOUBLE-STRANDED PROTEINS IN BACTERIA Russian patent published in 2019 - IPC A61K39/395 C07K16/24 C07K16/46 C12N15/70 

FIELD: biotechnology.

SUBSTANCE: group of inventions relates to biotechnology. Disclosed are methods of producing a polypeptide comprising two chains, in particular comprising a heavy chain and a light chain of a semi-antibody, particularly a semi-antibody against IL-13, as well as against IL-17 in a prokaryotic host cell, as well as methods for preparing a bispecific antibody containing a capable of binding a first antigen, in particular binding IL-13, a first semi-antibody and capable of binding a second antigen, particularly binding IL-17, a second semi-antibody. Methods include culturing a host cell comprising a polynucleotide for expressing two polypeptide chains at a growth phase temperature of 30 to 34 °C, temperature during phase of product formation from 25 to 29 °C and stirring speed at growth phase, at 50–250 rpm higher than the mixing rate during the product formation phase, and recovering the polypeptide, in particular a semi-antibody, from the host cell. Polynucleotide contains translation units which code two polypeptide chains, particularly antibodies, and at least one protein-chaperone. Methods of producing the bispecific antibody involve combining the obtained first semibody with a second semi-antibody.

EFFECT: invention ensures achievement of at least 10-fold increase of production of typical double-stranded polypeptides.

66 cl, 66 dwg, 7 tbl, 7 ex