Rapid, sensitive and quantitative methods for tissue and cell-based proteomics via consecutive addition of quantifiable extenders

A method for detecting and quantifying multiple immunogens in a sample comprising:

• (a) forming an immunogen-binding agent-DNA complex extended with two oligonucleotides, oligo 1 and oligo 2, by:
• (i) contacting an immobilized form of immunogens with an epitope detector comprising an immunogen-binding agent-DNA complex, wherein the DNA of the complex contains an RNA polymerase promoter;
• (ii) incubating the immobilized form of immunogens and the epitope detector under conditions which promote binding of the first immunogen-binding agent-DNA complex to an immunogen;
• (iii) ligating oligo 1 to the bound first immunogen-binding agent-DNA complex to form a complex immunogen-binding agent-DNA-oligo 1;
• (iv) ligating oligo 2 to the bound immunogen-binding agent-DNA-oligo 1 complex to form immunogen-binding agent-DNA-oligo 1-oligo 2;
• (b) repeating step (a) with different epitope detectors comprising Immunogen-binding-agents for different immunogens in the sample;
• (C) removing all bound immunogen-binding agent-DNA-oligo complexes from the immobilized immunogens;
• (d) adding RNA polymerase to the immunogen-binding agent-DNA-oligo complexes to form RNA products;
• and
• (e) analyzing RNA products wherein an amount of RNA product is directly proportional to the amount of immunogen-binding agent-DNA-oligo complexes bound to the immobilized immunogens in step (a) and size of an RNA product corresponds to the step at which the immunogen-binding agent was added with earlier added immunogen-binding agents giving rise to longer products while later added immunogen-binding agents produce smaller products.