Innovative production technique for antibody-enzyme

1 . A method for producing a κ light chain antibody having enzyme activity or improved enzyme activity, the method comprising:

a modification step comprising modifying a polynucleotide that encodes a κ light chain antibody having, as a variable region, a polypeptide having an amino acid sequence in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is a proline residue, by deleting or substituting the proline residue and obtaining a polynucleotide that encodes a κ light chain antibody having, as a variable region, a polypeptide having an amino acid sequence in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue; and

an expression step comprising expressing a κ light chain antibody having enzyme activity in an intracellular or extracellular expression system by using an expression vector including the polynucleotide that encodes a κ light chain antibody obtained after the modification step.

2 . The production method according to claim 1 ,

wherein the κ light chain antibody is a human κ light chain antibody or a mouse κ light chain antibody.

3 . The production method according to claim 1 ,

wherein a variable region of the κ light chain antibody having enzyme activity or improved enzyme activity is a polypeptide selected from the group consisting of the following (1a) to (3c),

(1a) a polypeptide having an amino acid sequence obtained by deletion or substitution of the 95th proline residue from the N-terminal of a variable region by the Kabat classification in an amino acid sequence represented by SEQ ID NO: 1;

(1b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence of the polypeptide (1a) and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(1c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence of the polypeptide (1a) and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(2a) an amino acid sequence obtained by deletion or substitution of the 95th proline residue from the N-terminal of a variable region by the Kabat classification in an amino acid sequence represented by SEQ ID NO: 2;

(2b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence of the polypeptide (2a) and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(2c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence of the polypeptide (2a) and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(3a) an amino acid sequence obtained by deletion or substitution of the 95th proline residue from the N-terminal of a variable region by the Kabat classification in an amino acid sequence represented by SEQ ID NO: 3;

(3b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence of the polypeptide (3a) and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(3c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence of the polypeptide (3a) and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue.

4 . The production method according to claim 1 ,

wherein a variable region of the κ light chain antibody having enzyme activity or improved enzyme activity is a polypeptide selected from the group consisting of the following (4a) to (7c),

(4a) a polypeptide having an amino acid sequence represented by SEQ ID NO: 4;

(4b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence represented by SEQ ID NO: 4 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(4c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence represented by SEQ ID NO: 4 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(5a) a polypeptide having an amino acid sequence represented by SEQ ID NO: 5;

(5b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence represented by SEQ ID NO: 5 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(5c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence represented by SEQ ID NO: 5 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(6a) a polypeptide having an amino acid sequence represented by SEQ ID NO: 6;

(6b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence represented by SEQ ID NO: 6 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(6c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence represented by SEQ ID NO: 6 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(7a) a polypeptide having an amino acid sequence represented by SEQ ID NO: 7;

(7b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence represented by SEQ ID NO: 7 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(7c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence represented by SEQ ID NO: 7 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue.

5 . The production method according to claim 1 ,

wherein a variable region of the κ light chain antibody having enzyme activity or improved enzyme activity is a polypeptide selected from the group consisting of the following (8a) to (8c),

(8a) an amino acid sequence obtained by deletion or substitution of the 95th proline residue from the N-terminal of a variable region by the Kabat classification in an amino acid sequence represented by SEQ ID NO: 8;

(8b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence of the polypeptide (8a) and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(8c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence of the polypeptide (8a) and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue.

6 . The production method according to claim 1 ,

wherein a variable region of the κ light chain antibody having enzyme activity is a polypeptide selected from the group consisting of the following (9a) to (9c),

(9a) a polypeptide having an amino acid sequence represented by SEQ ID NO: 9;

(9b) a polypeptide having an amino acid sequence which is obtained by substitution, addition, or deletion of one or more amino acids in the amino acid sequence represented by SEQ ID NO: 9 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue;

(9c) a polypeptide having an amino acid sequence which shares 90% or more identity with the amino acid sequence represented by SEQ ID NO: 9 and in which the 95th amino acid residue from the N-terminal of a variable region by the Kabat classification is deleted or substituted with an amino acid residue other than a proline residue.