Patent ID: 8658388

Claim:
A method for performing a multiplex, high-throughput immunoassay for facilitating a cancer diagnosis, the method comprising: (a) incubating a cellular extract with a plurality of dilution series of capture antibodies specific for at least one receptor tyrosine kinase in the cellular extract to form a plurality of captured receptor tyrosine kinases, wherein the capture antibodies are restrained on a solid support; (b) incubating the plurality of captured receptor tyrosine kinases with detection antibodies specific for the at least one receptor tyrosine kinase to form a plurality of detectable captured receptor tyrosine kinases, wherein the detection antibodies comprise: (1) a plurality of activation state-independent antibodies labeled with a facilitating moiety and specific for the at least one receptor tyrosine kinase, and (2) a plurality of activation state-dependent antibodies labeled with a first member of a signal amplification pair and specific for the at least one receptor tyrosine kinase, wherein the facilitating moiety, which is glucose oxidase conjugated to dextran, generates an oxidizing agent which channels to and reacts with the first member of the signal amplification pair, which is a peroxidase in proximity; (c) incubating the plurality of detectable captured receptor tyrosine kinases with a second member of the signal amplification pair to generate an amplified signal; (d) detecting the amplified signal generated from the first and second members of the signal amplification pair, and (e) measuring the activation state or level of the at least one receptor tyrosine kinases by monitoring the amplified signal, in the multiplex, high-throughput immunoassay for facilitating the cancer diagnosis, wherein the cancer is selected from the group consisting of lung cancer, digestive cancer, gastrointestinal cancer, esophageal cancer, gallbladder cancer, liver cancer, pancreatic cancer, appendix cancer, breast cancer, ovarian cancer, renal cancer, cancer of a central nervous system and a blood cancer.