METHODS FOR MAKING BISPECIFIC ANTIBODIES AGAINST CD3*CD19 IN FLEXYBODY FORMAT IN MAMMALIAN CELLS Russian patent published in 2016 - IPC C07K16/46 C07K16/22 C12P21/08 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. Method for producing a recombinant monoclonal antibody against CD3*CD19 in flexybody format. Constructed recombinant expression plasmid DNA encoding a bispecific antibody against the CD3*CD19. Resulting DNA is transfected with CHO cell strain M, and the resulting cell line is cultured. Cultivation is carried out in a perfusion mode in the presence of feeder additives CB1 to CB5 and 1: 1 ratio, with a volume of additive of 25 % by volume of the nutrient medium. Submitted monoclonal antibody against CD3*CD19 flexybody format having two polypeptide chains non-covalently associated with the amino acid sequence of SEQ ID №1 or non-covalently associated two chain polypeptide having the amino acid sequence of SEQ ID №2, obtained in this way. Also provided a recombinant expression plasmid DNA pOG776, encoding a polypeptide having the sequence bispecific antibody in flexybody format against CD19*CD3, having a size of 10,618 bp, and a recombinant expression plasmid DNA pOG854, encoding a polypeptide with the sequence of the bispecific anti-CD3 * CD19, having a size of 10,618 bp with the physical map shown in fig. 6.

EFFECT: invention provides a high yield of protein of pharmaceutical quality, suitable for the production of drugs.

6 cl, 8 dwg, 3 tbl, 6 ex