METHOD FOR DISPLAYING BISPECIFIC ANTIBODY ON MAMMALIAN CELL SURFACE AND VECTOR Russian patent published in 2025 - IPC A61K39/395 C07K16/18 

FIELD: biotechnology.

SUBSTANCE: obtaining a first polynucleotide containing S5-LC1-S6 in direction from 5' to 3'. Second polynucleotide containing B4-VH1-B3 in direction from 5' to 3' is obtained. Third polynucleotide containing B2-LC2-B4 in direction from 5' to 3' is obtained. Fourth polynucleotide containing B5-VH2-B6 in direction from 5' to 3' is obtained. Fifth polynucleotide containing S6-fragment I of expression vector-B2 in direction from 5' to 3' is obtained. Sixth polynucleotide containing B3-fragment II of expression vector-B5 in direction from 5' to 3' is obtained. Seventh polynucleotide containing the B6-fragment III of the expression vector-S5 in direction from 5' to 3' is obtained. First polynucleotide, the second polynucleotide, the third polynucleotide, the fourth polynucleotide, the fifth polynucleotide, the sixth polynucleotide and the seventh polynucleotide are specifically split using a restriction endonuclease. Restriction endonuclease specifically recognizes S5, S6, B4, B3, B2, B5 and B6, respectively. Split first, second, third, fourth, fifth, sixth and seventh split polynucleotide are mixed so that they can be ligated and cyclized to form an expression vector. LC1 codes the first light chain of the bispecific antigen-binding polypeptide, VH1 codes the variable region of the first heavy chain of the bispecific antigen-binding polypeptide, and the first light chain can bind to the variable region of the first heavy chain to form a first Fab for recognition of the first target; LC2 codes the second light chain of the bispecific antigen-binding polypeptide, VH2 codes the variable region of the second heavy chain of the bispecific antigen-binding polypeptide, and the second light chain can bind to the variable region of the second heavy chain to form a second Fab for recognition of the second target, and each of B2, B3, B4, B5, B6, S5 and S6 independently represent restriction endonuclease recognition sites.

EFFECT: described is a method for constructing an expression vector of a bispecific antigen-binding polypeptide.

16 cl, 4 dwg, 3 tbl, 3 ex