FIELD: bioengineering.
SUBSTANCE: antibody is suggested for use as bispecific antibody, characterized in that contains 4 polypeptides. The first and the second polypeptides contain: VH-domain linked with constant domain, and super helical domain. At the super helical domain is linked with the constant domain by decomposable linker, and contains in case of the first polypeptide the heptade repeat: (X1X2X3X4X5X6X7)n, and in case of the second polypeptide the super helical domain contains hepdate repeat: (X'1X'2X'3X'4X'5X'6X'7)n, where n is larger or equal to 2. X1, X'1 is hydrophobic amino-acid residue or asparagine, each of X2, X3, X6, X'2, X'3 and X'6 is any amino-acid residue, X4, X'4 is hydrophobic amino-acid residue, and each of X5, X7, X'5 and X'7 is charged amino-acid residue. At that in each heptade repeat the first super helical domain contains X5 residue that is opposite in terms of charge sign to X'7 residue of the second super helical domain, and the first super helical domain contains X7 residue that is opposite in terms of the charge sign to X'5 residue of the second super helical domain. The third and forth polypeptides contain the first and second VL-domains, respectively. Besides the antibody is suggested for use as heterodimeric one containing three polypeptides. At that the first polypeptide contains: VH-domain linked with constant domain, and super helical domain. The second polypeptide contains the constant domain containing CH2- and CH3-domains, and super helical domain. At that the super helical domain is linked with the constant domain by decomposed linker, and contains the heptade repeat same as in bispecific antibody. The third polypeptide contains VL-domain. Also the method of antibody production is described, it provides for stage of cell cultivation containing vector, coding the antibody under the invention in culture medium.
EFFECT: new method of antibodies dimerisation in presence of constant domains upon presence of the super helical domain, this can be used in production of the bispecific and heterodimeric antibodies.
17 cl, 32 dwg, 6 tbl, 7 ex
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Authors
Dates
2016-01-27—Published
2010-09-16—Filed