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US 10,501,737 B2Granted
Method for producing antigen-binding molecule using modified helper phage
AssigneeChugai Seiyaku Kabushiki Kaisha
InventorShinya Ishii
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Abstract
The present invention provides a method for preparing a bacteriophage displaying an antigen-binding molecule, comprising the step of contacting a helper phage capable of expressing a first polypeptide with a bacterium capable of expressing a second polypeptide, wherein the first polypeptide and the second polypeptide associate with each other to form the antigen-binding molecule.
Claims14
A method for preparing a plurality of multispecific antigen-binding molecules comprising common first polypeptides, wherein the method comprises:
- (a) carrying out for a plurality of different antigens a method for obtaining an antigen-binding molecule specifically binding to a predetermined antigen, wherein the method comprises the steps (a-1) and (a-2) below,
- (a-1) contacting the antigen with an antigen-binding molecule display library prepared by a method for preparing an antigen-binding molecule display library comprising common first polypeptides, wherein the method comprises the steps (1) and (2) below,
- (1) carrying out a plurality of times a method for preparing a bacteriophage displaying an antigen-binding molecule, comprising contacting a helper phage capable of expressing a first polypeptide with a bacterium capable of expressing a second polypeptide, wherein the first polypeptide and the second polypeptide associate with each other to form the antigen-binding molecule, wherein a plurality of bacteria used in the step are a bacterium population capable of expressing a plurality of second polypeptides differing in amino acid sequence, and helper phages used in the step are helper phages capable of expressing first polypeptides having identical amino acid sequences;
- and
- (2) recovering a plurality of bacteriophages displaying antigen-binding molecules prepared in (1);
- and
- (a-2) selecting an antigen-binding molecule binding to the antigen from the antigen-binding molecule display library;
- and
- (b) preparing a multispecific antigen-binding molecule using a plurality of the first polypeptides having identical amino acid sequences and a plurality of the second polypeptides having different amino acid sequences, contained in a plurality of antigen-binding molecules obtained in (a),
- wherein the first polypeptides associate with the plurality of second polypeptides, respectively, to form the plurality of antigen-binding sites specifically binding to the plurality of antigens.
7 more elements
A method for preparing a multispecific antigen-binding molecule comprising common first polypeptides, wherein the method comprises:
- (a) carrying out for a plurality of different antigens a method for obtaining an antigen-binding molecule specifically binding to a predetermined antigen, wherein the method comprises the steps (a-1) and (a-2) below,
- (a-1) contacting the antigen with an antigen-binding molecule display library prepared by a method for preparing an antigen-binding molecule display library comprising common first polypeptides, wherein the method comprises the steps (1) and (2) below,
- (1) carrying out a plurality of times a method for preparing a bacteriophage displaying an antigen-binding molecule, comprising contacting a helper phage capable of expressing a first polypeptide with a bacterium capable of expressing a second polypeptide, wherein the first polypeptide and the second polypeptide associate with each other to form the antigen-binding molecule, wherein a plurality of bacteria used in the step are a bacterium population capable of expressing a plurality of second polypeptides differing in amino acid sequence, and helper phages used in the step are helper phages capable of expressing first polypeptides having identical amino acid sequences;
- and
- (2) recovering a plurality of bacteriophages displaying antigen-binding molecules prepared in (1);
- and
- (a-2) selecting an antigen-binding molecule binding to the antigen from the antigen-binding molecule display library;
- (b) for a plurality of first polypeptides having identical amino acid sequences and a plurality of second polypeptides having different amino acid sequences, contained in a plurality of antigen-binding molecules obtained in (a), separately preparing polynucleotides encoding the first polypeptides and polynucleotides encoding the plurality of second polypeptides;
- (c) transferring each the polynucleotide prepared in (b) to a host cell;
- and
- (d) culturing the host cell of (c) to recover a multispecific antigen-binding molecule, wherein the first polypeptides associate with the plurality of second polypeptides, respectively, to form the plurality of antigen-binding sites specifically binding to the plurality of antigens.
8 more elements
Dependencies are inferred from the claim text.