Method for producing antibody-drug conjugate and enzyme used for the same

An enzyme agent for use in a reaction of a glycan-homogenized antibody with a glycosylated drug, the agent comprising any one of the following proteins:

• (a) a protein consisting of the amino acid sequence of SEQ ID NO:
• 1, or the amino acid sequence of SEQ ID NO:
• 1 in which the 165th amino acid is replaced with an amino acid other than aspartic acid (D);
• (b) a protein consisting of an amino acid sequence derived from the amino acid sequence of the protein described in (a) by substitution, deletion, insertion, and/or addition of one or more amino acids, provided that the 165th amino acid is an amino acid other than D, and having a conjugation activity;
• (c) a protein consisting of an amino acid sequence having an identity of at least 90% to the amino acid sequence of the protein described in (a), provided that the 165th amino acid is an amino acid other than D, and having a conjugation activity;
• (d) a protein consisting of the amino acid sequence of SEQ ID NO:
• 11, or the amino acid sequence of SEQ ID NO:
• 11 in which the 233rd amino acid is replaced with an amino acid other than D;
• (e) a protein consisting of an amino acid sequence derived from the amino acid sequence of the protein described in (d) by substitution, deletion, insertion, and/or addition of one or more amino acids, provided that the 233rd amino acid is an amino acid other than D, and having a conjugation activity;
• (f) protein consisting of an amino acid sequence having an identity of at least 90% to the amino acid sequence of the protein described in (d), provided that the 233rd amino acid is an amino acid other than D, and having a conjugation activity;
• and
• (g) a protein consisting of the amino acid sequence of SEQ ID NO:
• 24, or the amino acid sequence of SEQ ID NO:
• 24 in which the 165th amino acid is replaced with an amino acid other than D.

Use of a polynucleotide encoding any one of the following proteins in the production of an enzyme for use in a reaction of a glycan-homogenized antibody with a glycosylated drug:

• (a) a protein consisting of the amino acid sequence of SEQ ID NO:
• 1, or the amino acid sequence of SEQ ID NO:
• 1 in which the 165th amino acid is replaced with an amino acid other than aspartic acid (D);
• (b) a protein consisting of an amino acid sequence derived from the amino acid sequence of the protein described in (a) by substitution, deletion, insertion, and/or addition of one or more amino acids, provided that the 165th amino acid is an amino acid other than D, and having a conjugation activity;
• (c) a protein consisting of an amino acid sequence having an identity of at least 90% to the amino acid sequence of the protein described in (a), provided that the 165th amino acid is an amino acid other than D, and having a conjugation activity;
• (d) a protein consisting of the amino acid sequence of SEQ ID NO:
• 11, or the amino acid sequence of SEQ ID NO:
• 11 in which the 233rd amino acid is replaced with an amino acid other than D;
• (e) a protein consisting of an amino acid sequence derived from the amino acid sequence of the protein described in (d) by substitution, deletion, insertion, and/or addition of one or more amino acids, provided that the 233rd amino acid is an amino acid other than D, and having a conjugation activity;
• (f) a protein consisting of an amino acid sequence having an identity of at least 90% to the amino acid sequence of the protein described in (d), provided that the 233rd amino acid is an amino acid other than D, and having a conjugation activity;
• and
• (g) a protein consisting of the amino acid sequence of SEQ ID NO:
• 24, or the amino acid sequence of SEQ ID NO:
• 24 in which the 165th amino acid is replaced with an amino acid other than D.