Method for detecting bi-specific antibody complex
AssigneeChugai Seiyaku Kabushiki Kaisha
InventorsSotaro Naoi, Meiri Kawazoe
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Abstract
The present invention provides methods for detecting a complex with low affinity, under conditions in which the binding equilibrium of the complex is substantially maintained, and methods for measuring the concentration and/or amount of the complex. The invention also provides methods for evaluating the kinetics of a complex and methods for deciding on a therapeutic method that uses a pharmaceutical agent, based on the concentration and/or amount of the complex determined by the above-mentioned measurement method.
Claims11
A method for detecting a complex in a sample, the method comprising the steps of:
- (1) allowing a first binding entity to bind to a complex by bringing the first binding entity into contact with a sample comprising the complex;
- (2) allowing a second binding entity to bind to the complex bound to the first binding entity;
- and
- (3) detecting the second binding entity bound to the complex;
- wherein said complex comprises two or more components, at least any one of the KD values between the components is 1 nM or greater and at least two of the components are an antibody and an antigen recognized by the antibody, the first binding entity recognizes the antigen and is bound to a solid phase, and the method is conducted under a condition in which the time of contact between any particular complex in the sample and any particular antibody bound to the solid phase is ten seconds or less,
- wherein the first and second binding entities are antibodies, antibody fragments and fusion polypeptides comprising antibodies or antibody fragments thereof.
A method for detecting a complex in a sample, wherein the complex is a ternary complex comprising a bispecific antibody and a first antigen and second antigen recognized by the bispecific antibody as components, and wherein the method comprises the steps of:
- (1) allowing a first binding entity which recognizes the first antigen comprised in the complex to bind to the complex by bringing the first binding entity into contact with a sample containing the complex;
- (2) allowing a second binding entity which recognizes the second antigen to bind to the complex bound to the first binding entity;
- and
- (3) detecting the second binding entity bound to the complex
- wherein the first and second binding entities are antibodies, antibody fragments and fusion polypeptides comprising antibodies or antibody fragments thereof.
Dependencies are inferred from the claim text.