US 10,118,971 B2 · Protein in which electrical interaction is introduced within hydrophobic interaction site and preparation method therefor

Abstract

The present invention provides a protein or antibody in which, of a pair of hydrophobic amino acids selected from within a hydrophobic interaction site of the protein, one hydrophobic amino acid is transformed into a substance having a positive electrical charge and the other hydrophobic amino acid is transformed into a substance having a negative electrical charge, and electrostatic interaction is introduced within the hydrophobic interaction site of the protein by means of the positive charge and the negative charge. The present invention also provides a method for preparing the protein or antibody, and a method for measuring the degree of coupling between a heavy chain and a light chain, using the antibody. The protein or antibody in accordance with to the present invention has a low contamination by a homodimer or a monomer, and thus a heterodimer can be obtained in high purity.

Claims9

Dependent claims

A method of preparation of a protein with increased chain selectivity, the method comprising the following steps:

(a) selecting at least two pairs of hydrophobic amino acids from the group consisting of 351 leucine and 351 leucine pair, 395 proline and 397 valine pair, and 395 proline and 395 proline pair between CH3 domains in a protein comprising two Fc domains of human IgG1 of SEQ ID NO:
1, or IgG4 of SEQ ID NO:
4;
(b) modifying one hydrophobic amino acid to a positive charged amino acid selected from the group consisting of lysine, arginine and histidine, and the other hydrophobic amino acid to a negative charged amino acid selected from the group consisting of aspartic acid and glutamic acid, in the selected pairs of hydrophobic amino acids;
and
(c) binding with electrostatic interaction by contacting between the positive charged amino acid and the negative charged amino acid.

3 more elements

A protein comprising two Fc domains of human IgG1 of SEQ ID NO:

1, or IgG4 of SEQ ID NO:
4, characterized in that at least two hydrophobic amino acid pairs selected from the group consisting of 351 leucine and 351 leucine pair, 395 proline and 397 valine pair, and 395 proline and 395 proline pair between CH3 domains of the protein are mutated by substituting one hydrophobic amino acid of each amino acid pair with a positive charged amino acid selected from the group consisting of lysine, arginine and histidine, and the other hydrophobic amino acid with a negative charged amino acid selected from the group consisting of aspartic acid and glutamic acid, whereby electrostatic interaction is introduced between the mutated amino acids in the protein.

Dependencies are inferred from the claim text.

Protein in which electrical interaction is introduced within hydrophobic interaction site and preparation method therefor

Life of this patent

Protein in which electrical interaction is introduced within hydrophobic interaction site and preparation method therefor

Life of this patent

Abstract

Claims9