JP2016502091A5

JP2016502091A5 -

Info

Publication number: JP2016502091A5
Application number: JP2015544592A
Authority: JP
Filing date: 2013-11-27
Publication date: 2017-01-05

Claims

A method for detecting a neutralized form of an autoantibody against an anti-TNFα drug in a sample comprising:
(A) said sample is contacted with a labeled anti-TNF [alpha] agents and labeled TNF [alpha],
(I) forming said labeled first labeled complex of anti-TNFα agents and said autoantibodies, and / or (ii) the second labeled complex of the labeled anti-TNFα agents and the labeled TNFα and the autoantibodies When,
(B) said first labeled complex and / or the second labeled complex was subjected to size exclusion chromatography, the first labeled complex and / or the second labeled complex, free label TNF [alpha], free label Separating from the anti-TNFα drug and / or the complex of labeled anti-TNFα drug and labeled TNFα;
Measuring the (c) the free peak area under the curve of labeled TNFα in said sample after size exclusion chromatography (AUC),
Step (d) the AUC of the peak of said measured free label TNFα in (c), by comparing the AUC of the peak of the free label TNFα in a control sample, the autoantibodies against the anti-TNFα agents in the sample Detecting the neutralized form of the control sample, wherein the control sample is a reference sample to which only free labeled TNFα has been added; and
Including methods.

It said anti-TNFα agents, Lee Nfurikishimabu, et Taneruseputo, A Darimumabu, Se belt Liz Mab Bae goal, Gore Rimuma Bed (CNTO 148), and is selected from the group consisting of combinations thereof The method of claim 1.

The autoantibodies against the anti-TNFα agents, human anti-chimeric antibody (HACA), human anti-humanized antibodies (HAHA), is selected from the group consisting of human anti-mouse antibody (HAMA), and combinations thereof, according to claim 1 or 2. The method according to 2.

AUC of the peak of the free label TNFα of the control sample, is measured after size exclusion chromatography, The method according to any one of claims 1 to 3.

It said sample is serum, the method according to any one of claims 1-4.

Said sample, said a sample from a subject undergoing treatment with an anti-TNFα agents, the method according to any one of claims 1 to 5.

Comprising said labeled anti-TNFα agents and the labeled TNFα is different fluorophores or fluorescent dyes A method according to any one of claims 1 to 6.

The method according to any one of claims 1 to 7 , wherein the neutralized form of the autoantibodies is present in a population of autoantibodies comprising neutralizing autoantibodies and non-neutralizing autoantibodies.

9. The method according to any one of claims 1 to 8, wherein the reference sample is normal human serum (NHS).

A method for determining whether a neutralized form of an autoantibody against a first anti-TNFα drug is cross-reactive with a second anti-TNFα drug, comprising:
Neutralization shape of said autoantibodies in (a) sample, is detected by performing a method according to any one of claims 1-9, wherein the sample is positive for neutralization forms of the autoantibody Determining whether it is negative or negative,
If the sample for neutralization forms of the autoantibodies are positive,
A step (b) said sample is contacted with labeled second anti-TNFα agents, to form a third labeled complex with a neutralizing form of the autoantibody and the labeled second anti-TNFα agents,
(C) subjecting the third labeled complex to size exclusion chromatography to separate the third labeled complex;
(D) detecting the third labeled complex, thereby determining whether the neutralized form of the autoantibody against the first anti-TNFα drug is cross-reactive with the second anti-TNFα drug;
A method further comprising:

It said first and second anti-TNFα drugs, Lee Nfurikishimabu, et Taneruseputo, A Darimumabu, Se belt Liz Mab Bae goal, Gore Rimuma Bed (CNTO 148), and are independently selected from the group consisting a combination thereof, wherein Item 11. The method according to Item 10 .

Wherein said autoantibodies against first anti TNFα drugs, human anti-chimeric antibody (HACA), human anti-humanized antibodies (HAHA), is selected from the group consisting of human anti-mouse antibody (HAMA), and combinations thereof, according to claim The method according to 10 or 11 .

The presence of the third labeled complex indicates that neutralizing autoantibodies against the first anti-TNFα agent is cross-reactive with the second anti-TNFα drugs, according to any one of claims 10 to 12 the method of.

Absence of the third labeled complex indicates that neutralizing autoantibodies against the first anti-TNFα agents is not cross-reactive with the second anti-TNFα drugs, according to any one of claims 10 to 12 the method of.

The method according to claim 10 , wherein the sample is serum.

Said sample, said a sample from a subject undergoing treatment with an anti-TNFα agents, the method according to any one of claims 10 to 15.

The method according to any one of claims 10 to 16 , wherein the labeled second anti-TNFα drug comprises a phosphor or a fluorescent dye.

In a subject undergoing treatment course with anti-TNFα agents, the monitoring of the treatment of anti-TNFα agents or a method for optimizing the treatment of the anti-TNFα agents,
Neutralization form of autoantibodies against (a) the anti-TNFα agents, by carrying out the method according to any one of claims 1 to 9, at multiple time points during the course of treatment, a step of detecting,
(B) detecting the temporal change neutralization shaped percent or level of the autoantibody,
Subsequent doses of treatment course for (c) the subject, or whether to be carried out on the subject to different treatment course, determined based on the temporal change percentage or level of neutralizing type of said autoantibodies Steps,
Including methods.

A method for optimizing treatment and / or reducing toxicity in a subject undergoing a course of treatment with a first anti-TNFα drug comprising:
Neutralization form of autoantibody in a sample from (a) the subject, by detecting by performing a method according to any one of claims 1 to 9, the autoantibodies against the first anti-TNFα agents Determining whether the neutralized form is cross-reactive with a second anti-TNFα drug;
(B) if the neutralization form of the autoantibody is cross-reactive with the second anti-TNFα agents, and determining the to implement the different treatment courses to the target,
Including methods.

A method for determining whether a sample is positive for a neutralized form of an autoantibody against an anti-TNFα drug comprising:
(A) contacting the sample with a labeled anti-TNFα drug and labeled TNFα;
(I) a first labeled complex of the labeled anti-TNFα drug and the autoantibody, and / or
(Ii) a second labeled complex of the labeled anti-TNFα drug, the labeled TNFα and the autoantibody
Forming a step;
(B) subjecting the first labeled complex and / or the second labeled complex to size exclusion chromatography to convert the first labeled complex and / or the second labeled complex into free labeled TNFα and free labeled Separating from the anti-TNFα drug and / or the complex of labeled anti-TNFα drug and labeled TNFα;
(C) Area under the curve (AUC) of the peak of the free labeled TNFα in the sample after size exclusion chromatography (free labeled TNFα of the sample _ＡＵＣAUC ) Measuring,
(D) AUC of the peak of free labeled TNFα in the control sample (free labeled TNFα of the control sample _ＡＵＣAUC ), The percent of neutralized form of the autoantibody is expressed by the following formula:
(Free labeled TNFα of sample _ＡＵＣAUC -Background) / (Free labeled TNFα of control sample) _ＡＵＣAUC -Background) * 100,
The control sample is a reference sample containing only free labeled TNFα, and
(E) if the percentage of the neutralized form of the autoantibody calculated in step (d) is greater than a threshold of about 1.25% to about 1.30%, the sample is about the neutralized form of the autoantibody Determining to be positive,
Including a method.