A method of detecting magea4
OwnerAmanda WILLIAMSinventor
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Abstract
The present invention provides a method of detecting MAGEA4 in a sample. The method may comprise adding an anti-MAGEA4 antibody to the sample at a concentration in the range of from 2-20 μg/ml, incubating the antibody and the sample and detecting antibody that is bound to the sample. The methods of the invention can be used to diagnose whether a subject has cancer and whether the subject is eligible for treatment with a MAGEA4 targeted cancer therapy.
Claims20
A method of detecting MAGEA4 in a sample, the method comprising:
- adding an anti-MAGEA4 antibody to the sample at a concentration in the range of from 2-20 μg/ml;
- incubating the antibody and the sample;
- and
- detecting antibody bound to the sample, wherein the antibody is OTI1F9.
The method of any preceding claim, wherein the sample and the anti-MAGEA4 antibody are incubated for 30-60, 25-50, 25-39, 27-35, 28-34, 30-33, 31, 32 or 33 minutes and/or at 33-39, 34-38, 35-37, 35, 36 or 37° C.
The method of any preceding claim, wherein the sample and the anti-MAGEA4 antibody are incubated for 32 minutes at 36° C.
The method of any preceding claim, wherein anti-MAGEA4 antibody bound to the sample is detected indirectly by a secondary antibody that binds to the anti-MAGEA4 antibody.
The method of any preceding claim, wherein the secondary antibody is conjugated to horseradish peroxidase.
The method of any preceding claim, further comprising the step of epitope retrieval prior to adding the anti-MAGEA4 antibody to the sample.
The method of any preceding claim, further comprising inhibiting endogenous peroxidase activity in the sample, optionally by contacting the sample with a DAB (3,3′-Diaminobenzidine) inhibitor.
The method of any preceding claim, further comprising the step of contacting the sample with a blocking reagent, optionally after inhibiting endogenous peroxidase activity.
The method of any preceding claim, wherein the sample is embedded in paraffin
The method of any preceding claim, comprising:
- deparaffinising the sample, optionally by incubating the sample in reaction buffer such as EZ-Prep solution, optionally for 3 cycles of 8 minutes at 69° C.;
- retrieving epitope, optionally by incubating the sample in cell conditioning fluid 1 at 95° C. for 20 mins;
- inhibiting endogenous peroxidase activity, optionally by incubating the sample in DAB inhibitor for 8 mins at room temperature;
- incubating the sample in blocking reagent, optionally for 12 mins at 36° C.;
- adding antibody OFI1F9 to the sample at a concentration of 10 μg/ml;
- incubating the antibody and the sample for 32 mins at 36° C.;
- and
- detecting antibody that is bound to the sample.
Dependencies are inferred from the claim text.