Patent ID: 9726669
Date: 2017-08-08
CPC Classifications: G01N

Claim:
1. A method for determining the sensitivity of a test cell to a compound that modulates HER2 activity, the method comprising: (a) contacting the test cell with the compound; (b) lysing the test cell to produce a cellular extract; (c) determining the activation level of HER2 and p95HER2 in the cellular extract produced from the test cell using a proximity immunoassay comprising capture antibodies, activation state-dependent antibodies, and activation state-independent antibodies, wherein the activation state-independent antibodies are labeled with a facilitating moiety, the activation state-dependent antibodies are labeled with a first member of a signal amplification pair, and the facilitating moiety generates an oxidizing agent which channels to and reacts with the first member of the signal amplification pair, thereby allowing the first member of the signal amplification pair to generate an amplified signal when incubated with a second member of the signal amplification pair; (d) determining a first reference activation level of HER2 and p95HER2 in a cellular extract produced from a compound-sensitive cell that is treated with the compound and a second reference activation level of HER2 and p95HER2 in a cellular extract produced from a compound-resistant cell that is treated with the compound using the proximity immunoassay, wherein the level of p95HER2 activation in the compound-resistant cell is at least 5-fold higher than the level of p95HER2 activation in the compound-sensitive cell; and (e) determining that the test cell is sensitive to the compound if the activation level of HER2 and p95HER2 in the cellular extract produced from the test cell is equal to or at least about 1.5-fold lower compared to the first reference activation level of HER2 and p95HER2 or